LOCUS       sequence-271741-        7164 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expression of ZONAB isoform A in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7164)
  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 7164)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-A
FEATURES             Location/Qualifiers
     source          1..7164
                     /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    1606..2228
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1805..1824)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2252..2272)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2270..2567
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2621..3415
                     /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(2675..2694)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3285..3304
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4067..4201
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4104..4123)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4158..4177
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4206..4225)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4207..4225)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4239..4255)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4239..4255)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4252..4274)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4263..4279
                     /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(4287..4293,4294..4311,4312..4317))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4332..4353
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4470..4487)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4641..5229)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4721..4740)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5400..6191,6192..6260))
                     /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     6023..6042
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6261..6365)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6392..6847
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6479..6498)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6689..6710
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6973..6995
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6987..7004
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6988..7004
                     /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 atgccggtga gattggagag atgaaggagg gagtcccaga ggggacacag ctgcagggac
     1321 cggttcatcg aaatccaact taccgcccaa ggtaccggag aggccctcct cgcccccgac
     1381 ctgccccagc tgttggtgag gctgaagata aagagaacca acaagcagcc aatggtccaa
     1441 accagccatc tgctcgccgt ggataccggc gcccatacaa ctatcggcgt cgcccccgcc
     1501 cccctaatgc tccttcacaa gatggcaaag agaccaaggc aggtgaagtt ccatctgaga
     1561 accccgctcc agccaccgag cagagcagtg ctgagtaagg atcccgggtg gcatccctgt
     1621 gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt
     1681 gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta taatattatg
     1741 gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc
     1801 ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc
     1861 gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg attccaggca
     1921 tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg
     1981 gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg
     2041 ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttctgatttt aaaataacta
     2101 taccagcagg aggacgtcca gacacagcat aggctacctg gccatgccca accggtggga
     2161 catttgagtt gcttgcttgg cactgtcctc tcatgcgttg ggtccactca gtagatgcct
     2221 gttgaattgg gtacgcggcc agcttggctg tggaatgtgt gtcagttagg gtgtggaaag
     2281 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     2341 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     2401 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     2461 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     2521 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg aattgatcag
     2581 cttgggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     2641 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     2701 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     2761 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     2821 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     2881 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     2941 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     3001 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     3061 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     3121 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     3181 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     3241 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     3301 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     3361 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     3421 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     3481 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     3541 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccgg gctcgatccc
     3601 ctcgcgagtt ggttcagctg ctgcctgagg ctggacgacc tcgcggagtt ctaccggcag
     3661 tgcaaatccg tcggcatcca ggaaaccagc agcggctatc cgcgcatcca tgcccccgaa
     3721 ctgcaggagt ggggaggcac gatggccgct ttggtcgacc cggacgggac gctcctgcgc
     3781 ctgatacaga acgaattgct tgcaggcatc tcatgagtgt gtcttcccgt tttccgcctg
     3841 aggtcactgc gtggatggag cgctggcgcc tgctgcgcga cggcgagctg ctcaccaccc
     3901 actcgccaag ctggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     3961 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     4021 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     4081 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4141 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat
     4201 caattcccta tagtgagtcg tattaaattc gtaatcatgt catagctgtt tcctgtgtga
     4261 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     4321 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     4381 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     4441 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     4501 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4561 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4621 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4681 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4741 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4801 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4861 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4921 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4981 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5041 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5101 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5161 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5221 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5281 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     5341 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     5401 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     5461 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     5521 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     5581 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     5641 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     5701 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     5761 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     5821 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     5881 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     5941 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6001 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6061 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6121 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6181 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6241 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6301 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6361 ccgcgcacat ttccccgaaa agtgccacct gacgcgccct gtagcggcgc attaagcgcg
     6421 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     6481 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     6541 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     6601 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     6661 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     6721 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     6781 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     6841 acaatttcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     6901 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     6961 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgccaag ctgatctata
     7021 cattgaatca atattggcaa ttagccatat tagtcattgg ttatatagca taaatcaata
     7081 ttggctattg gccattgcat acgttgtatc tatatcataa tatgtacatt tatattggct
     7141 catgtccaat atgaccgcca tgtt
//
