LOCUS       sequence-271733-        7368 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ZONAB isoform B in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7368)
  AUTHORS   Balda MS, Matter K
  TITLE     The tight junction protein ZO-1 and an interacting transcription 
            factor regulate ErbB-2 expression.
  JOURNAL   EMBO J. 2000 May 2;19(9):2024-33. doi: 10.1093/emboj/19.9.2024.
  PUBMED    10790369
REFERENCE   2  (bases 1 to 7368)
  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: 
            "10.1093/emboj/19.9"; journalName: "EMBO J"; date: "2000-05-2- 2"; 
            volume: "19"; issue: "9"; pages: "2024-33"
COMMENT     Sequence Label: pCB6-ZONAB-B
FEATURES             Location/Qualifiers
     source          1..7368
                     /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    1810..2432
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2009..2028)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2456..2476)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2474..2771
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2825..3619
                     /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(2879..2898)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3489..3508
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4271..4405
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4308..4327)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4362..4381
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4410..4429)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4411..4429)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4443..4459)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4443..4459)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4456..4478)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4467..4483
                     /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(4491..4497,4498..4515,4516..4521))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4536..4557
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4674..4691)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4845..5433)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4925..4944)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5604..6395,6396..6464))
                     /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     6227..6246
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6465..6569)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6596..7051
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6683..6702)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6893..6914
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7177..7199
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7191..7208
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7192..7208
                     /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 gtaccacgcg tatcgataag cttgcatgcc
      661 tgcaggtcga ctctagagga tccatgagcg aggcgggcga ggccaccggc accaccagca
      721 gcagcagcag cagcctcccg caggctccgg cggaggcggc ccccgcggcg ccccaggacc
      781 ccgcgcccaa gagcccgggg ggcagcggcg cgccccaggc cgcggccccg ccgcccgccg
      841 cgctcgccgc gggaaacccc ggtggggacg cggcccccgc agccacgggc accgcggccc
      901 ccgcctcttc ggcccccgcc ggcggtgagg acgcggagaa gaaagttctc gccaccaaag
      961 tccttggcac tgtcaaatgg ttcaacgtca gaaatggata tggatttata aatcgaaatg
     1021 acaccaaaga agatgtattt gtacatcaga ctgccatcaa gaagaataac ccacggaaat
     1081 acctgcgcag tgtaggagat ggagaaactg tagaatttga tgtggttgaa ggagagaagg
     1141 gtgcagaagc agccaatgtg actggaccag acggagttcc tgtagaaggg agccgttacg
     1201 ctgctgatcg gcgtcgatac agacgtggtt actatggcag acgccgtgga cctccccgta
     1261 attacgctgg ggaggaggag gaggaaggga gcggcagcag tgaaggattt gacccccctg
     1321 ccactgacgg gcagttctct ggggcccgga gtcagctgcg ccgcccccag tatcgccctc
     1381 agtaccggag gcggttcccg ccttaccacg tgggacagac ctttgaccgt cgctcacggg
     1441 tctttcccca tcccaacaga atgcaggccg gtgagattgg agagatgaag gagggagtcc
     1501 cagaggggac acagctgcag ggaccggttc atcgaaatcc aacttaccgc ccaaggtacc
     1561 ggagaggccc tcctcgcccc cgacctgccc cagctgttgg tgaggctgaa gataaagaga
     1621 accaacaagc agccaatggt ccaaaccagc catctgctcg ccgtggatac cggcgcccat
     1681 acaactatcg gcgtcgcccc cgccccccta atgctccttc acaagatggc aaagagacca
     1741 aggcaggtga agttccatct gagaaccccg ctccagccac cgagcagagc agtgctgagt
     1801 aaggatcccg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc cctggaagtt
     1861 gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat tttgtctgac
     1921 taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa ggggcaagtt
     1981 gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag tgcagtggca
     2041 caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct gcctcagcct
     2101 cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt gtttttttgg
     2161 tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc tcaggtgatc
     2221 tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc tcccttccct
     2281 gtccttctga ttttaaaata actataccag caggaggacg tccagacaca gcataggcta
     2341 cctggccatg cccaaccggt gggacatttg agttgcttgc ttggcactgt cctctcatgc
     2401 gttgggtcca ctcagtagat gcctgttgaa ttgggtacgc ggccagcttg gctgtggaat
     2461 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     2521 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     2581 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     2641 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     2701 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     2761 ggcttttttg gaggaattga tcagcttggg atctgatcaa gagacaggat gaggatcgtt
     2821 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     2881 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     2941 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     3001 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     3061 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     3121 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     3181 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     3241 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     3301 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     3361 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     3421 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     3481 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     3541 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     3601 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     3661 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     3721 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     3781 ttcgcccacc ccgggctcga tcccctcgcg agttggttca gctgctgcct gaggctggac
     3841 gacctcgcgg agttctaccg gcagtgcaaa tccgtcggca tccaggaaac cagcagcggc
     3901 tatccgcgca tccatgcccc cgaactgcag gagtggggag gcacgatggc cgctttggtc
     3961 gacccggacg ggacgctcct gcgcctgata cagaacgaat tgcttgcagg catctcatga
     4021 gtgtgtcttc ccgttttccg cctgaggtca ctgcgtggat ggagcgctgg cgcctgctgc
     4081 gcgacggcga gctgctcacc acccactcgc caagctggaa ccgtaaaaag gccgcgttgc
     4141 tggcgttttt ccataggctc cgccgatcat aatcagccat accacatttg tagaggtttt
     4201 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     4261 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4321 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     4381 caatgtatct tatcatgtct ggatcaattc cctatagtga gtcgtattaa attcgtaatc
     4441 atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     4501 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     4561 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     4621 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     4681 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     4741 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     4801 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4861 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     4921 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4981 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     5041 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5101 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5161 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5221 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5281 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5341 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5401 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     5461 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     5521 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     5581 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     5641 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     5701 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     5761 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     5821 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     5881 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     5941 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6001 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6061 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6121 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6181 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6241 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6301 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6361 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     6421 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     6481 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6541 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg
     6601 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     6661 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     6721 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     6781 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     6841 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     6901 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     6961 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     7021 aattttaaca aaatattaac gcttacaatt tccattcgcc attcaggctg cgcaactgtt
     7081 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     7141 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     7201 cggccagtgc caagctgatc tatacattga atcaatattg gcaattagcc atattagtca
     7261 ttggttatat agcataaatc aatattggct attggccatt gcatacgttg tatctatatc
     7321 ataatatgta catttatatt ggctcatgtc caatatgacc gccatgtt
//
