LOCUS       sequence-82721-e        8999 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of nuclear LacZ driven by Gfa2 promoter with 3 additional
            B sites, has about 75 times higher activity than pGfa2-nLac in 
            transfected cells .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8999)
  AUTHORS   de Leeuw B, Su M, ter Horst M, Iwata S, Rodijk M, Hoeben RC, Messing
            A, Smitt PS, Brenner M
  TITLE     Increased glia-specific transgene expression with glial fibrillary 
            acidic protein promoters containing multiple enhancer elements.
  JOURNAL   J Neurosci Res. 2006 Apr;83(5):744-53.
  PUBMED    16496373
REFERENCE   2  (bases 1 to 8999)
  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"; journalName: "J Neurosci 
            Res."; date: "2006-04"; volume: "83(5)"; pages: "744-53"
COMMENT     Sequence Label: pGfa(B3)-nLac
FEATURES             Location/Qualifiers
     source          1..8999
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    106..127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(142..147,148..165,166..172)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /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)."
     primer_bind     185..207
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     204..220
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2920..2940
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2959..6006
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase"
                     /label=lacZ
                     /translation="VVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRS
                     LNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNVTYPITVNPP
                     FVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYGQDSRLPSEF
                     DLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQISDFHVATR
                     FNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIIDERGGYADRV
                     TLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIENGLLLLNGKP
                     LLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLWYTLCDRYGL
                     YVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWSLGNESGHGA
                     NHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVPKWSIKKWLS
                     LPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSLIKYDENGNP
                     WSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQTIEVTSEYL
                     FRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQLWLTVRVVQ
                     PNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELGNKRWQFNRQ
                     SGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWKAAGHYQAEA
                     ALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDVEVASDTPHP
                     ARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPYVFPSENGLR
                     CGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDGFHMGIGGDD
                     SWSPSVSAEFQLSAGRYHYQLVWCQK"
     primer_bind     complement(6603..6620)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6603..6619)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6828..6847)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6947..6969
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7007..7025)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7093..7197
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7198..7266,7267..8058)
                     /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"
     rep_origin      8229..8817
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8718..8737
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8971..8988
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg aattcgagct
      241 cggtaccaga tctgagctcc cacctccctc tctgtgctgg gactcacaga gggagacctc
      301 aggaggcagt ctgtccatca catgtccaaa tgcagagcat accctgggct gggcgcagtg
      361 gcgcacaact gtaattccag cactttggga ggctgatgtg gaaggatcac ttgagcccag
      421 aagttctaga ccagcctggg caacatggca agaccctatc tctacaaaaa aagttaaaaa
      481 atcagccacg tgtggtgaca cacacctgta gtcccagcta ttcaggaggc tgaggtgagg
      541 ggatcactta aggctgggag gttgaggctg cagtgagtcg tggttgcgcc actgcactcc
      601 agcctgggca acagtgagac cctgtctcaa aagacaaaaa aaaaaaaaaa aaaaaaaaga
      661 acatatcctg gtgtggagta ggggacgctg ctctgacaga ggctcggggg cctgagctgg
      721 ctctgtgagc tggggaggag gcagacagcc aggccttgtc tgcaagcaga cctggcagca
      781 ttgggctggc cgccccccag ggcctcctct tcatgcccag tgaatgactc accttggcac
      841 agacacaatg ttcggggtgg gcacagtgcc tgcttcccgc cgcaccccag cccccctcaa
      901 atgccttccg agaagcccat tgagcagggg gcttgcattg caccccagcc tgacagcctg
      961 gcatcttggg ataaaagcag cacagccccc taggggctgc ccttgctgtg tggcgccacc
     1021 ggcggtggag aacaaggctc tattcagcct gtgcccagga aaggggatca ggggatgccc
     1081 aggcatggac agtgggtggc agggggggag aggagggctg tctgcttccc agaagtccaa
     1141 ggacacaaat gggtgagggg actgggcagg gttctgaccc tgtgggacca gagtggaggg
     1201 cgtagatgga cctgaagtct ccagggacaa cagggcccag gtctcaggct cctagttggg
     1261 cccagtggct ccagcgtttc caaacccatc catccccaga ggttcttccc atctctccag
     1321 gctgatgtgt gggaactcga ggaaataaat ctccagtggg agacggaggg gtggccaggg
     1381 aaacggggcg ctgcaggaat aaagacgagc cagcacagcc agctcatgtg taacggcttt
     1441 gtggagctgt caaggcctgg tctctgggag agaggcacag ggaggccaga caaggaaggg
     1501 gtgacctgga gggacagatc caggggctaa agtcctgata aggcaagaga gtgccggccc
     1561 cctcttgccc tatcaggacc tccactgcca catagaggcc atgattgacc cttagacaaa
     1621 gggctggtgt ccaatcccag cccccagccc cagaactcca gggaatgaat gggcagagag
     1681 caggaatgtg ggacatctgt gttcaaggga aggactccag gagtctgctg ggaatgaggc
     1741 ctagtaggaa atgaggtggc ccttgagggt acagaacagg ttcattcttc gccaaattcc
     1801 cagcaccttg caggcactta cagctgagtg agataatgcc tgggttatga aatcaaaaag
     1861 ttggaaagca ggtcagaggt catctggtac agcccttcct tccctttttt tttttttttt
     1921 tttgtgagac aaggtctctc tctgttgccc aggctggagt ggcgcaaaca cagctcactg
     1981 cagcctcaac ctactgggct caagcaatcc tccagcctca gcctcccaaa gtgctgggat
     2041 tacaagcatg agccacccca ctcagccctt tccttccttt ttaattgatg cataataatt
     2101 gtaagtattc atcatggtcc aaccaaccct ttcttgaccc accttcctag agagagggtc
     2161 ctcttgcttc agcggtcagg gccccagacc catggtctgg ctccaggtac cacctgcctc
     2221 atgcaggagt tggcgtgccc aggaagctct gcctctgggc acagtgacct cagtggggtg
     2281 aggggagctc tccccatagc tgggctgcgg cccaacccca ccccctcagg ctatgccagg
     2341 gggtgttgcc aggggcaccc gatcttcctc ttcatgccca gtgaatgact caccttggca
     2401 cagacacaat gttcggggtg ggcacagtgc ctgcttcccg ccgcacccca gcccccctca
     2461 aatgccttcc gagaagccca ttgagcaggc ccgatcttcc tcttcatgcc cagtgaatga
     2521 ctcaccttgg cacagacaca atgttcgggg tgggcacagt gcctgcttcc cgccgcaccc
     2581 cagcccccct caaatgcctt ccgagaagcc cattgagcag gcccgatctt cctcttcatg
     2641 cccagtgaat gactcacctt ggcacagaca caatgttcgg ggtgggcaca gtgcctgctt
     2701 cccgccgcac cccagccccc ctcaaatgcc ttccgagaag cccattgagc aggcccgggc
     2761 atcgccagtc tagcccactc cttcataaag ccctcgcatc ccaggagcga gcagagccag
     2821 agcaggttgg agaggagacg catcacctcc gctgctcgcg gggatcctct agagtcgacg
     2881 gatccgggga attccccagt ctcaggatcc accatggggc ccaagaagaa acgcaaagtg
     2941 gggagcatgg gggatcccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     3001 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     3061 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg ctttgcctgg
     3121 tttccggcac cagaagcggt gccggaaagc tggctggagt gcgatcttcc tgaggccgat
     3181 actgtcgtcg tcccctcaaa ctggcagatg cacggttacg atgcgcccat ctacaccaac
     3241 gtaacctatc ccattacggt caatccgccg tttgttccca cggagaatcc gacgggttgt
     3301 tactcgctca catttaatgt tgatgaaagc tggctacagg aaggccagac gcgaattatt
     3361 tttgatggcg ttaactcggc gtttcatctg tggtgcaacg ggcgctgggt cggttacggc
     3421 caggacagtc gtttgccgtc tgaatttgac ctgagcgcat ttttacgcgc cggagaaaac
     3481 cgcctcgcgg tgatggtgct gcgttggagt gacggcagtt atctggaaga tcaggatatg
     3541 tggcggatga gcggcatttt ccgtgacgtc tcgttgctgc ataaaccgac tacacaaatc
     3601 agcgatttcc atgttgccac tcgctttaat gatgatttca gccgcgctgt actggaggct
     3661 gaagttcaga tgtgcggcga gttgcgtgac tacctacggg taacagtttc tttatggcag
     3721 ggtgaaacgc aggtcgccag cggcaccgcg cctttcggcg gtgaaattat cgatgagcgt
     3781 ggtggttatg ccgatcgcgt cacactacgt ctgaacgtcg aaaacccgaa actgtggagc
     3841 gccgaaatcc cgaatctcta tcgtgcggtg gttgaactgc acaccgccga cggcacgctg
     3901 attgaagcag aagcctgcga tgtcggtttc cgcgaggtgc ggattgaaaa tggtctgctg
     3961 ctgctgaacg gcaagccgtt gctgattcga ggcgttaacc gtcacgagca tcatcctctg
     4021 catggtcagg tcatggatga gcagacgatg gtgcaggata tcctgctgat gaagcagaac
     4081 aactttaacg ccgtgcgctg ttcgcattat ccgaaccatc cgctgtggta cacgctgtgc
     4141 gaccgctacg gcctgtatgt ggtggatgaa gccaatattg aaacccacgg catggtgcca
     4201 atgaatcgtc tgaccgatga tccgcgctgg ctaccggcga tgagcgaacg cgtaacgcga
     4261 atggtgcagc gcgatcgtaa tcacccgagt gtgatcatct ggtcgctggg gaatgaatca
     4321 ggccacggcg ctaatcacga cgcgctgtat cgctggatca aatctgtcga tccttcccgc
     4381 ccggtgcagt atgaaggcgg cggagccgac accacggcca ccgatattat ttgcccgatg
     4441 tacgcgcgcg tggatgaaga ccagcccttc ccggctgtgc cgaaatggtc catcaaaaaa
     4501 tggctttcgc tacctggaga gacgcgcccg ctgatccttt gcgaatacgc ccacgcgatg
     4561 ggtaacagtc ttggcggttt cgctaaatac tggcaggcgt ttcgtcagta tccccgttta
     4621 cagggcggct tcgtctggga ctgggtggat cagtcgctga ttaaatatga tgaaaacggc
     4681 aacccgtggt cggcttacgg cggtgatttt ggcgatacgc cgaacgatcg ccagttctgt
     4741 atgaacggtc tggtctttgc cgaccgcacg ccgcatccag cgctgacgga agcaaaacac
     4801 cagcagcagt ttttccagtt ccgtttatcc gggcaaacca tcgaagtgac cagcgaatac
     4861 ctgttccgtc atagcgataa cgagctcctg cactggatgg tggcgctgga tggtaagccg
     4921 ctggcaagcg gtgaagtgcc tctggatgtc gctccacaag gtaaacagtt gattgaactg
     4981 cctgaactac cgcagccgga gagcgccggg caactctggc tcacagtacg cgtagtgcaa
     5041 ccgaacgcga ccgcatggtc agaagccggg cacatcagcg cctggcagca gtggcgtctg
     5101 gcggaaaacc tcagtgtgac gctccccgcc gcgtcccacg ccatcccgca tctgaccacc
     5161 agcgaaatgg atttttgcat cgagctgggt aataagcgtt ggcaatttaa ccgccagtca
     5221 ggctttcttt cacagatgtg gattggcgat aaaaaacaac tgctgacgcc gctgcgcgat
     5281 cagttcaccc gtgcaccgct ggataacgac attggcgtaa gtgaagcgac ccgcattgac
     5341 cctaacgcct gggtcgaacg ctggaaggcg gcgggccatt accaggccga agcagcgttg
     5401 ttgcagtgca cggcagatac acttgctgat gcggtgctga ttacgaccgc tcacgcgtgg
     5461 cagcatcagg ggaaaacctt atttatcagc cggaaaacct accggattga tggtagtggt
     5521 caaatggcga ttaccgttga tgttgaagtg gcgagcgata caccgcatcc ggcgcggatt
     5581 ggcctgaact gccagctggc gcaggtagca gagcgggtaa actggctcgg attagggccg
     5641 caagaaaact atcccgaccg ccttactgcc gcctgttttg accgctggga tctgccattg
     5701 tcagacatgt ataccccgta cgtcttcccg agcgaaaacg gtctgcgctg cgggacgcgc
     5761 gaattgaatt atggcccaca ccagtggcgc ggcgacttcc agttcaacat cagccgctac
     5821 agtcaacagc aactgatgga aaccagccat cgccatctgc tgcacgcgga agaaggcaca
     5881 tggctgaata tcgacggttt ccatatgggg attggtggcg acgactcctg gagcccgtca
     5941 gtatcggcgg aattccagct gagcgccggt cgctaccatt accagttggt ctggtgtcaa
     6001 aaataataat aaccgggcag gggggatccg cagatcccgg ccagataccg atgctgccgc
     6061 agcaaaagca ggagcagatg ccgccgtcgc aggcgaagat gtcgcagacg gaggaggcga
     6121 tgctgccggc ggaggaggcg aagtaagtag agggctgggc tgggctgtgg ggggtgtggg
     6181 gtgcgggact gggcagtctg ggagtccctc tcaccacttt tcttaccttt ctaggatgct
     6241 gccgtcgccg ccgctcatac accataaggt gtaaaaaata ctagatgcac agaatagcaa
     6301 gtccatcaaa actcctgcgt gagaatttta ccagacttca agagcatctc gccacatctt
     6361 gaaaaatgcc accgtccgat gaaaaacagg agcctgctaa ggaacaatgc cacctgtcaa
     6421 taaatgttga aaactcatcc cattcctgcc tcttggtcct tgggcttggg gaggggtgcg
     6481 cggatgtggt tagggaacat gactggtcaa atgggaaggg cttcaaaaga attcccaata
     6541 ttgactacca agccacctgt acagatcgaa ttcagatctg cctgcaggca tgcaagcttg
     6601 gcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     6661 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     6721 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc
     6781 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct
     6841 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     6901 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     6961 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     7021 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     7081 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     7141 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     7201 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     7261 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     7321 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     7381 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     7441 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     7501 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     7561 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     7621 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     7681 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     7741 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     7801 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     7861 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     7921 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     7981 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     8041 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     8101 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     8161 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     8221 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     8281 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     8341 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     8401 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     8461 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     8521 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     8581 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     8641 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     8701 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     8761 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     8821 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     8881 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     8941 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaaga
//
