LOCUS       sequence-82730-e        6829 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of LacZ driven by partial human gfa2 promoter, expression
            is localized to a subset of astrocytes in mice. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6829)
  AUTHORS   Lee Y, Su M, Messing A, Brenner M
  TITLE     Astrocyte heterogeneity revealed by expression of a GFAP-LacZ 
            transgene.
  JOURNAL   Glia. 2006 May;53(7):677-87.
  PUBMED    16482522
REFERENCE   2  (bases 1 to 6829)
  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: "Glia."; 
            date: "2006-05"; volume: "53(7)"; pages: "677-87"
COMMENT     Sequence Label: pGfa28-nLac
FEATURES             Location/Qualifiers
     source          1..6829
                     /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             761..781
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             800..3847
                     /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(4433..4450)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4433..4449)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4658..4677)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4777..4799
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4837..4855)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4923..5027
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5028..5096,5097..5888)
                     /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     complement(5246..5265)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6059..6647
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6548..6567
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6801..6818
                     /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 atgattacga attcgagctc
      241 ggtaccagat ctaacatatc ctggtgtgga gtaggggacg ctgctctgac agaggctcgg
      301 gggcctgagc tggctctgtg agctggggag gaggcagaca gccaggcctt gtctgcaagc
      361 agacctggca gcattgggct ggccgccccc cagggcctcc tcttcatgcc cagtgaatga
      421 ctcaccttgg cacagacaca atgttcgggg tgggcacagt gcctgcttcc cgccgcaccc
      481 cagcccccct caaatgcctt ccgagaagcc cattgagcag ggagctctcc ccatagctgg
      541 gctgcggccc aaccccaccc cctcaggcta tgccaggggg tgttgccagg ggcacccggg
      601 catcgccagt ctagcccact ccttcataaa gccctcgcat cccaggagcg agcagagcca
      661 gagcaggttg gagaggagac gcatcacctc cgctgctcgc ggggatcctc tagagtcgac
      721 ggatccgggg aattccccag tctcaggatc caccatgggg cccaagaaga aacgcaaagt
      781 ggggagcatg ggggatcccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
      841 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
      901 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gctttgcctg
      961 gtttccggca ccagaagcgg tgccggaaag ctggctggag tgcgatcttc ctgaggccga
     1021 tactgtcgtc gtcccctcaa actggcagat gcacggttac gatgcgccca tctacaccaa
     1081 cgtaacctat cccattacgg tcaatccgcc gtttgttccc acggagaatc cgacgggttg
     1141 ttactcgctc acatttaatg ttgatgaaag ctggctacag gaaggccaga cgcgaattat
     1201 ttttgatggc gttaactcgg cgtttcatct gtggtgcaac gggcgctggg tcggttacgg
     1261 ccaggacagt cgtttgccgt ctgaatttga cctgagcgca tttttacgcg ccggagaaaa
     1321 ccgcctcgcg gtgatggtgc tgcgttggag tgacggcagt tatctggaag atcaggatat
     1381 gtggcggatg agcggcattt tccgtgacgt ctcgttgctg cataaaccga ctacacaaat
     1441 cagcgatttc catgttgcca ctcgctttaa tgatgatttc agccgcgctg tactggaggc
     1501 tgaagttcag atgtgcggcg agttgcgtga ctacctacgg gtaacagttt ctttatggca
     1561 gggtgaaacg caggtcgcca gcggcaccgc gcctttcggc ggtgaaatta tcgatgagcg
     1621 tggtggttat gccgatcgcg tcacactacg tctgaacgtc gaaaacccga aactgtggag
     1681 cgccgaaatc ccgaatctct atcgtgcggt ggttgaactg cacaccgccg acggcacgct
     1741 gattgaagca gaagcctgcg atgtcggttt ccgcgaggtg cggattgaaa atggtctgct
     1801 gctgctgaac ggcaagccgt tgctgattcg aggcgttaac cgtcacgagc atcatcctct
     1861 gcatggtcag gtcatggatg agcagacgat ggtgcaggat atcctgctga tgaagcagaa
     1921 caactttaac gccgtgcgct gttcgcatta tccgaaccat ccgctgtggt acacgctgtg
     1981 cgaccgctac ggcctgtatg tggtggatga agccaatatt gaaacccacg gcatggtgcc
     2041 aatgaatcgt ctgaccgatg atccgcgctg gctaccggcg atgagcgaac gcgtaacgcg
     2101 aatggtgcag cgcgatcgta atcacccgag tgtgatcatc tggtcgctgg ggaatgaatc
     2161 aggccacggc gctaatcacg acgcgctgta tcgctggatc aaatctgtcg atccttcccg
     2221 cccggtgcag tatgaaggcg gcggagccga caccacggcc accgatatta tttgcccgat
     2281 gtacgcgcgc gtggatgaag accagccctt cccggctgtg ccgaaatggt ccatcaaaaa
     2341 atggctttcg ctacctggag agacgcgccc gctgatcctt tgcgaatacg cccacgcgat
     2401 gggtaacagt cttggcggtt tcgctaaata ctggcaggcg tttcgtcagt atccccgttt
     2461 acagggcggc ttcgtctggg actgggtgga tcagtcgctg attaaatatg atgaaaacgg
     2521 caacccgtgg tcggcttacg gcggtgattt tggcgatacg ccgaacgatc gccagttctg
     2581 tatgaacggt ctggtctttg ccgaccgcac gccgcatcca gcgctgacgg aagcaaaaca
     2641 ccagcagcag tttttccagt tccgtttatc cgggcaaacc atcgaagtga ccagcgaata
     2701 cctgttccgt catagcgata acgagctcct gcactggatg gtggcgctgg atggtaagcc
     2761 gctggcaagc ggtgaagtgc ctctggatgt cgctccacaa ggtaaacagt tgattgaact
     2821 gcctgaacta ccgcagccgg agagcgccgg gcaactctgg ctcacagtac gcgtagtgca
     2881 accgaacgcg accgcatggt cagaagccgg gcacatcagc gcctggcagc agtggcgtct
     2941 ggcggaaaac ctcagtgtga cgctccccgc cgcgtcccac gccatcccgc atctgaccac
     3001 cagcgaaatg gatttttgca tcgagctggg taataagcgt tggcaattta accgccagtc
     3061 aggctttctt tcacagatgt ggattggcga taaaaaacaa ctgctgacgc cgctgcgcga
     3121 tcagttcacc cgtgcaccgc tggataacga cattggcgta agtgaagcga cccgcattga
     3181 ccctaacgcc tgggtcgaac gctggaaggc ggcgggccat taccaggccg aagcagcgtt
     3241 gttgcagtgc acggcagata cacttgctga tgcggtgctg attacgaccg ctcacgcgtg
     3301 gcagcatcag gggaaaacct tatttatcag ccggaaaacc taccggattg atggtagtgg
     3361 tcaaatggcg attaccgttg atgttgaagt ggcgagcgat acaccgcatc cggcgcggat
     3421 tggcctgaac tgccagctgg cgcaggtagc agagcgggta aactggctcg gattagggcc
     3481 gcaagaaaac tatcccgacc gccttactgc cgcctgtttt gaccgctggg atctgccatt
     3541 gtcagacatg tataccccgt acgtcttccc gagcgaaaac ggtctgcgct gcgggacgcg
     3601 cgaattgaat tatggcccac accagtggcg cggcgacttc cagttcaaca tcagccgcta
     3661 cagtcaacag caactgatgg aaaccagcca tcgccatctg ctgcacgcgg aagaaggcac
     3721 atggctgaat atcgacggtt tccatatggg gattggtggc gacgactcct ggagcccgtc
     3781 agtatcggcg gaattccagc tgagcgccgg tcgctaccat taccagttgg tctggtgtca
     3841 aaaataataa taaccgggca ggggggatcc gcagatcccg gccagatacc gatgctgccg
     3901 cagcaaaagc aggagcagat gccgccgtcg caggcgaaga tgtcgcagac ggaggaggcg
     3961 atgctgccgg cggaggaggc gaagtaagta gagggctggg ctgggctgtg gggggtgtgg
     4021 ggtgcgggac tgggcagtct gggagtccct ctcaccactt ttcttacctt tctaggatgc
     4081 tgccgtcgcc gccgctcata caccataagg tgtaaaaaat actagatgca cagaatagca
     4141 agtccatcaa aactcctgcg tgagaatttt accagacttc aagagcatct cgccacatct
     4201 tgaaaaatgc caccgtccga tgaaaaacag gagcctgcta aggaacaatg ccacctgtca
     4261 ataaatgttg aaaactcatc ccattcctgc ctcttggtcc ttgggcttgg ggaggggtgc
     4321 gcggatgtgg ttagggaaca tgactggtca aatgggaagg gcttcaaaag aattcccaat
     4381 attgactacc aagccacctg tacagatctg cctgcaggca tgcaagcttg gcactggccg
     4441 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     4501 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     4561 aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc cttacgcatc
     4621 tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat
     4681 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     4741 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     4801 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     4861 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     4921 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4981 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     5041 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     5101 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     5161 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     5221 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     5281 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     5341 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     5401 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     5461 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     5521 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     5581 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     5641 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     5701 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     5761 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     5821 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     5881 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     5941 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     6001 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     6061 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     6121 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     6181 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     6241 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     6301 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     6361 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     6421 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     6481 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     6541 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     6601 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6661 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     6721 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     6781 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaaga
//
