LOCUS pGfa2-nLac 8591 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Expression of LacZ driven by the human Gfa2 promoter. LacZ can also
be replaced with the gene of interest for targeting transgene
expression to astrocytes in mice..
ACCESSION .
VERSION .
KEYWORDS pGfa2-nLac
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8591)
AUTHORS Brenner M, Kisseberth WC, Su Y, Besnard F, Messing A
TITLE GFAP promoter directs astrocyte-specific expression in transgenic
mice.
JOURNAL J Neurosci. 1994 Mar;14(3 Pt 1):1030-7.
PUBMED 8120611
REFERENCE 2 (bases 1 to 8591)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8591
/organism="synthetic DNA construct"
/mol_type="other DNA"
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 142..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 2523..2543
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 2562..5609
/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(6195..6212)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(6195..6211)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6420..6439)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 6539..6561
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(6599..6617)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 6685..6789
/gene="bla"
/label=AmpR promoter
CDS 6790..7650
/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(7008..7027)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 7821..8409
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 8310..8329
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 8563..8580
/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 ctgagctccc acctccctct ctgtgctggg actcacagag ggagacctca
301 ggaggcagtc tgtccatcac atgtccaaat gcagagcata ccctgggctg ggcgcagtgg
361 cgcacaactg taattccagc actttgggag gctgatgtgg aaggatcact tgagcccaga
421 agttctagac cagcctgggc aacatggcaa gaccctatct ctacaaaaaa agttaaaaaa
481 tcagccacgt gtggtgacac acacctgtag tcccagctat tcaggaggct gaggtgaggg
541 gatcacttaa ggctgggagg ttgaggctgc agtgagtcgt ggttgcgcca ctgcactcca
601 gcctgggcaa cagtgagacc ctgtctcaaa agacaaaaaa aaaaaaaaaa aaaaaaagaa
661 catatcctgg tgtggagtag gggacgctgc tctgacagag gctcgggggc ctgagctggc
721 tctgtgagct ggggaggagg cagacagcca ggccttgtct gcaagcagac ctggcagcat
781 tgggctggcc gccccccagg gcctcctctt catgcccagt gaatgactca ccttggcaca
841 gacacaatgt tcggggtggg cacagtgcct gcttcccgcc gcaccccagc ccccctcaaa
901 tgccttccga gaagcccatt gagcaggggg cttgcattgc accccagcct gacagcctgg
961 catcttggga taaaagcagc acagccccct aggggctgcc cttgctgtgt ggcgccaccg
1021 gcggtggaga acaaggctct attcagcctg tgcccaggaa aggggatcag gggatgccca
1081 ggcatggaca gtgggtggca gggggggaga ggagggctgt ctgcttccca gaagtccaag
1141 gacacaaatg ggtgagggga ctgggcaggg ttctgaccct gtgggaccag agtggagggc
1201 gtagatggac ctgaagtctc cagggacaac agggcccagg tctcaggctc ctagttgggc
1261 ccagtggctc cagcgtttcc aaacccatcc atccccagag gttcttccca tctctccagg
1321 ctgatgtgtg ggaactcgag gaaataaatc tccagtggga gacggagggg tggccaggga
1381 aacggggcgc tgcaggaata aagacgagcc agcacagcca gctcatgtgt aacggctttg
1441 tggagctgtc aaggcctggt ctctgggaga gaggcacagg gaggccagac aaggaagggg
1501 tgacctggag ggacagatcc aggggctaaa gtcctgataa ggcaagagag tgccggcccc
1561 ctcttgccct atcaggacct ccactgccac atagaggcca tgattgaccc ttagacaaag
1621 ggctggtgtc caatcccagc ccccagcccc agaactccag ggaatgaatg ggcagagagc
1681 aggaatgtgg gacatctgtg ttcaagggaa ggactccagg agtctgctgg gaatgaggcc
1741 tagtaggaaa tgaggtggcc cttgagggta cagaacaggt tcattcttcg ccaaattccc
1801 agcaccttgc aggcacttac agctgagtga gataatgcct gggttatgaa atcaaaaagt
1861 tggaaagcag gtcagaggtc atctggtaca gcccttcctt cccttttttt tttttttttt
1921 ttgtgagaca aggtctctct ctgttgccca ggctggagtg gcgcaaacac agctcactgc
1981 agcctcaacc tactgggctc aagcaatcct ccagcctcag cctcccaaag tgctgggatt
2041 acaagcatga gccaccccac tcagcccttt ccttcctttt taattgatgc ataataattg
2101 taagtattca tcatggtcca accaaccctt tcttgaccca ccttcctaga gagagggtcc
2161 tcttgcttca gcggtcaggg ccccagaccc atggtctggc tccaggtacc acctgcctca
2221 tgcaggagtt ggcgtgccca ggaagctctg cctctgggca cagtgacctc agtggggtga
2281 ggggagctct ccccatagct gggctgcggc ccaaccccac cccctcaggc tatgccaggg
2341 ggtgttgcca ggggcacccg ggcatcgcca gtctagccca ctccttcata aagccctcgc
2401 atcccaggag cgagcagagc cagagcaggt tggagaggag acgcatcacc tccgctgctc
2461 gcggggatcc tctagagtcg acggatccgg ggaattcccc agtctcagga tccaccatgg
2521 ggcccaagaa gaaacgcaaa gtggggagca tgggggatcc cgtcgtttta caacgtcgtg
2581 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
2641 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
2701 atggcgaatg gcgctttgcc tggtttccgg caccagaagc ggtgccggaa agctggctgg
2761 agtgcgatct tcctgaggcc gatactgtcg tcgtcccctc aaactggcag atgcacggtt
2821 acgatgcgcc catctacacc aacgtaacct atcccattac ggtcaatccg ccgtttgttc
2881 ccacggagaa tccgacgggt tgttactcgc tcacatttaa tgttgatgaa agctggctac
2941 aggaaggcca gacgcgaatt atttttgatg gcgttaactc ggcgtttcat ctgtggtgca
3001 acgggcgctg ggtcggttac ggccaggaca gtcgtttgcc gtctgaattt gacctgagcg
3061 catttttacg cgccggagaa aaccgcctcg cggtgatggt gctgcgttgg agtgacggca
3121 gttatctgga agatcaggat atgtggcgga tgagcggcat tttccgtgac gtctcgttgc
3181 tgcataaacc gactacacaa atcagcgatt tccatgttgc cactcgcttt aatgatgatt
3241 tcagccgcgc tgtactggag gctgaagttc agatgtgcgg cgagttgcgt gactacctac
3301 gggtaacagt ttctttatgg cagggtgaaa cgcaggtcgc cagcggcacc gcgcctttcg
3361 gcggtgaaat tatcgatgag cgtggtggtt atgccgatcg cgtcacacta cgtctgaacg
3421 tcgaaaaccc gaaactgtgg agcgccgaaa tcccgaatct ctatcgtgcg gtggttgaac
3481 tgcacaccgc cgacggcacg ctgattgaag cagaagcctg cgatgtcggt ttccgcgagg
3541 tgcggattga aaatggtctg ctgctgctga acggcaagcc gttgctgatt cgaggcgtta
3601 accgtcacga gcatcatcct ctgcatggtc aggtcatgga tgagcagacg atggtgcagg
3661 atatcctgct gatgaagcag aacaacttta acgccgtgcg ctgttcgcat tatccgaacc
3721 atccgctgtg gtacacgctg tgcgaccgct acggcctgta tgtggtggat gaagccaata
3781 ttgaaaccca cggcatggtg ccaatgaatc gtctgaccga tgatccgcgc tggctaccgg
3841 cgatgagcga acgcgtaacg cgaatggtgc agcgcgatcg taatcacccg agtgtgatca
3901 tctggtcgct ggggaatgaa tcaggccacg gcgctaatca cgacgcgctg tatcgctgga
3961 tcaaatctgt cgatccttcc cgcccggtgc agtatgaagg cggcggagcc gacaccacgg
4021 ccaccgatat tatttgcccg atgtacgcgc gcgtggatga agaccagccc ttcccggctg
4081 tgccgaaatg gtccatcaaa aaatggcttt cgctacctgg agagacgcgc ccgctgatcc
4141 tttgcgaata cgcccacgcg atgggtaaca gtcttggcgg tttcgctaaa tactggcagg
4201 cgtttcgtca gtatccccgt ttacagggcg gcttcgtctg ggactgggtg gatcagtcgc
4261 tgattaaata tgatgaaaac ggcaacccgt ggtcggctta cggcggtgat tttggcgata
4321 cgccgaacga tcgccagttc tgtatgaacg gtctggtctt tgccgaccgc acgccgcatc
4381 cagcgctgac ggaagcaaaa caccagcagc agtttttcca gttccgttta tccgggcaaa
4441 ccatcgaagt gaccagcgaa tacctgttcc gtcatagcga taacgagctc ctgcactgga
4501 tggtggcgct ggatggtaag ccgctggcaa gcggtgaagt gcctctggat gtcgctccac
4561 aaggtaaaca gttgattgaa ctgcctgaac taccgcagcc ggagagcgcc gggcaactct
4621 ggctcacagt acgcgtagtg caaccgaacg cgaccgcatg gtcagaagcc gggcacatca
4681 gcgcctggca gcagtggcgt ctggcggaaa acctcagtgt gacgctcccc gccgcgtccc
4741 acgccatccc gcatctgacc accagcgaaa tggatttttg catcgagctg ggtaataagc
4801 gttggcaatt taaccgccag tcaggctttc tttcacagat gtggattggc gataaaaaac
4861 aactgctgac gccgctgcgc gatcagttca cccgtgcacc gctggataac gacattggcg
4921 taagtgaagc gacccgcatt gaccctaacg cctgggtcga acgctggaag gcggcgggcc
4981 attaccaggc cgaagcagcg ttgttgcagt gcacggcaga tacacttgct gatgcggtgc
5041 tgattacgac cgctcacgcg tggcagcatc aggggaaaac cttatttatc agccggaaaa
5101 cctaccggat tgatggtagt ggtcaaatgg cgattaccgt tgatgttgaa gtggcgagcg
5161 atacaccgca tccggcgcgg attggcctga actgccagct ggcgcaggta gcagagcggg
5221 taaactggct cggattaggg ccgcaagaaa actatcccga ccgccttact gccgcctgtt
5281 ttgaccgctg ggatctgcca ttgtcagaca tgtatacccc gtacgtcttc ccgagcgaaa
5341 acggtctgcg ctgcgggacg cgcgaattga attatggccc acaccagtgg cgcggcgact
5401 tccagttcaa catcagccgc tacagtcaac agcaactgat ggaaaccagc catcgccatc
5461 tgctgcacgc ggaagaaggc acatggctga atatcgacgg tttccatatg gggattggtg
5521 gcgacgactc ctggagcccg tcagtatcgg cggaattcca gctgagcgcc ggtcgctacc
5581 attaccagtt ggtctggtgt caaaaataat aataaccggg caggggggat ccgcagatcc
5641 cggccagata ccgatgctgc cgcagcaaaa gcaggagcag atgccgccgt cgcaggcgaa
5701 gatgtcgcag acggaggagg cgatgctgcc ggcggaggag gcgaagtaag tagagggctg
5761 ggctgggctg tggggggtgt ggggtgcggg actgggcagt ctgggagtcc ctctcaccac
5821 ttttcttacc tttctaggat gctgccgtcg ccgccgctca tacaccataa ggtgtaaaaa
5881 atactagatg cacagaatag caagtccatc aaaactcctg cgtgagaatt ttaccagact
5941 tcaagagcat ctcgccacat cttgaaaaat gccaccgtcc gatgaaaaac aggagcctgc
6001 taaggaacaa tgccacctgt caataaatgt tgaaaactca tcccattcct gcctcttggt
6061 ccttgggctt ggggaggggt gcgcggatgt ggttagggaa catgactggt caaatgggaa
6121 gggcttcaaa agaattccca atattgacta ccaagccacc tgtacagatc tgcctgcagg
6181 catgcaagct tggcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
6241 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
6301 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg
6361 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt
6421 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
6481 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
6541 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
6601 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
6661 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
6721 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
6781 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
6841 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
6901 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
6961 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
7021 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
7081 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
7141 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
7201 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
7261 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
7321 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
7381 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
7441 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
7501 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
7561 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
7621 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
7681 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
7741 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
7801 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
7861 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
7921 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
7981 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
8041 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
8101 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
8161 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
8221 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
8281 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
8341 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
8401 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
8461 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
8521 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
8581 gaagcggaag a
//
