LOCUS       sequence-263310-        7062 bp    DNA     circular SYN 14-APR-2025
DEFINITION  expression of lacZ driven by human GfaABC1D promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7062)
  AUTHORS   Lee Y, Messing A, Su M, Brenner M
  TITLE     GFAP promoter elements required for region-specific and 
            astrocyte-specific expression.
  JOURNAL   Glia. 2008 Apr;56(5):481-93. doi: 10.1002/glia.20622.
  PUBMED    18240313
REFERENCE   2  (bases 1 to 7062)
  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: "10.1002/glia"; 
            journalName: "Glia"; date: "2008-04"; volume: "56"; issue: "5"; 
            pages: "481-93"
COMMENT     Sequence Label: pGfaABC1D-nLac
FEATURES             Location/Qualifiers
     source          1..7062
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(13..31)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        99..203
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(204..272,273..1064)
                     /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      1235..1823
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1724..1743
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1977..1994
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2111..2132
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2147..2152,2153..2170,2171..2177)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2185..2201
                     /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     2190..2212
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2209..2225
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2209..2225
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2999..3019
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3038..6085
                     /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(6671..6688)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6671..6687)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6896..6915)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7015..7037
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
       61 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
      121 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
      181 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
      241 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
      301 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
      361 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
      421 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
      481 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
      541 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
      601 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
      661 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
      721 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
      781 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
      841 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
      901 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
      961 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     1021 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     1081 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     1141 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     1201 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     1261 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     1321 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     1381 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     1441 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     1501 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     1561 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     1621 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     1681 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     1741 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     1801 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     1861 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     1921 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     1981 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     2041 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     2101 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
     2161 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
     2221 atgacatgat tacgaattcg agctcggtac cagatctaac atatcctggt gtggagtagg
     2281 ggacgctgct ctgacagagg ctcgggggcc tgagctggct ctgtgagctg gggaggaggc
     2341 agacagccag gccttgtctg caagcagacc tggcagcatt gggctggccg ccccccaggg
     2401 cctcctcttc atgcccagtg aatgactcac cttggcacag acacaatgtt cggggtgggc
     2461 acagtgcctg cttcccgccg caccccagcc cccctcaaat gccttccgag aagcccattg
     2521 agcagggggc ttgcattgca ccccagcctg acagcctggc atcttgggat aaaagcagca
     2581 cagcccccta ggggctgccc ttgctgtgtg gcgccaccgg cggtggagaa caaggctcta
     2641 ttcagcctgt gcccaggaaa ggggatcagg ggatgcccag gcatggacag tgggtggcag
     2701 ggggggagag gagggctgtc tgcttcccag aagtccaagg acacaaatgg gtgaggggag
     2761 agctctcccc atagctgggc tgcggcccaa ccccaccccc tcaggctatg ccagggggtg
     2821 ttgccagggg cacccgggca tcgccagtct agcccactcc ttcataaagc cctcgcatcc
     2881 caggagcgag cagagccaga gcaggttgga gaggagacgc atcacctccg ctgctcgcgg
     2941 ggatcctcta gagtcgacgg atccggggaa ttccccagtc tcaggatcca ccatggggcc
     3001 caagaagaaa cgcaaagtgg ggagcatggg ggatcccgtc gttttacaac gtcgtgactg
     3061 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     3121 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     3181 cgaatggcgc tttgcctggt ttccggcacc agaagcggtg ccggaaagct ggctggagtg
     3241 cgatcttcct gaggccgata ctgtcgtcgt cccctcaaac tggcagatgc acggttacga
     3301 tgcgcccatc tacaccaacg tgacctatcc cattacggtc aatccgccgt ttgttcccac
     3361 ggagaatccg acgggttgtt actcgctcac atttaatgtt gatgaaagct ggctacagga
     3421 aggccagacg cgaattattt ttgatggcgt taactcggcg tttcatctgt ggtgcaacgg
     3481 gcgctgggtc ggttacggcc aggacagtcg tttgccgtct gaatttgacc tgagcgcatt
     3541 tttacgcgcc ggagaaaacc gcctcgcggt gatggtgctg cgctggagtg acggcagtta
     3601 tctggaagat caggatatgt ggcggatgag cggcattttc cgtgacgtct cgttgctgca
     3661 taaaccgact acacaaatca gcgatttcca tgttgccact cgctttaatg atgatttcag
     3721 ccgcgctgta ctggaggctg aagttcagat gtgcggcgag ttgcgtgact acctacgggt
     3781 aacagtttct ttatggcagg gtgaaacgca ggtcgccagc ggcaccgcgc ctttcggcgg
     3841 tgaaattatc gatgagcgtg gtggttatgc cgatcgcgtc acactacgtc tgaacgtcga
     3901 aaacccgaaa ctgtggagcg ccgaaatccc gaatctctat cgtgcggtgg ttgaactgca
     3961 caccgccgac ggcacgctga ttgaagcaga agcctgcgat gtcggtttcc gcgaggtgcg
     4021 gattgaaaat ggtctgctgc tgctgaacgg caagccgttg ctgattcgag gcgttaaccg
     4081 tcacgagcat catcctctgc atggtcaggt catggatgag cagacgatgg tgcaggatat
     4141 cctgctgatg aagcagaaca actttaacgc cgtgcgctgt tcgcattatc cgaaccatcc
     4201 gctgtggtac acgctgtgcg accgctacgg cctgtatgtg gtggatgaag ccaatattga
     4261 aacccacggc atggtgccaa tgaatcgtct gaccgatgat ccgcgctggc taccggcgat
     4321 gagcgaacgc gtaacgcgaa tggtgcagcg cgatcgtaat cacccgagtg tgatcatctg
     4381 gtcgctgggg aatgaatcag gccacggcgc taatcacgac gcgctgtatc gctggatcaa
     4441 atctgtcgat ccttcccgcc cggtgcagta tgaaggcggc ggagccgaca ccacggccac
     4501 cgatattatt tgcccgatgt acgcgcgcgt ggatgaagac cagcccttcc cggctgtgcc
     4561 gaaatggtcc atcaaaaaat ggctttcgct acctggagag acgcgcccgc tgatcctttg
     4621 cgaatacgcc cacgcgatgg gtaacagtct tggcggtttc gctaaatact ggcaggcgtt
     4681 tcgtcagtat ccccgtttac agggcggctt cgtctgggac tgggtggatc agtcgctgat
     4741 taaatatgat gaaaacggca acccgtggtc ggcttacggc ggtgattttg gcgatacgcc
     4801 gaacgatcgc cagttctgta tgaacggtct ggtctttgcc gaccgcacgc cgcatccagc
     4861 gctgacggaa gcaaaacacc agcagcagtt tttccagttc cgtttatccg ggcaaaccat
     4921 cgaagtgacc agcgaatacc tgttccgtca tagcgataac gagctcctgc actggatggt
     4981 ggcgctggat ggtaagccgc tggcaagcgg tgaagtgcct ctggatgtcg ctccacaagg
     5041 taaacagttg attgaactgc ctgaactacc gcagccggag agcgccgggc aactctggct
     5101 cacagtacgc gtagtgcaac cgaacgcgac cgcatggtca gaagccgggc acatcagcgc
     5161 ctggcagcag tggcgtctgg cggaaaacct cagtgtgacg ctccccgccg cgtcccacgc
     5221 catcccgcat ctgaccacca gcgaaatgga tttttgcatc gagctgggta ataagcgttg
     5281 gcaatttaac cgccagtcag gctttctttc acagatgtgg attggcgata aaaaacaact
     5341 gctgacgccg ctgcgcgatc agttcacccg tgcaccgctg gataacgaca ttggcgtaag
     5401 tgaagcgacc cgcattgacc ctaacgcctg ggtcgaacgc tggaaggcgg cgggccatta
     5461 ccaggccgaa gcagcgttgt tgcagtgcac ggcagataca cttgctgatg cggtgctgat
     5521 tacgaccgct cacgcgtggc agcatcaggg gaaaacctta tttatcagcc ggaaaaccta
     5581 ccggattgat ggtagtggtc aaatggcgat taccgttgat gttgaagtgg cgagcgatac
     5641 accgcatccg gcgcggattg gcctgaactg ccagctggcg caggtagcag agcgggtaaa
     5701 ctggctcgga ttagggccgc aagaaaacta tcccgaccgc cttactgccg cctgttttga
     5761 ccgctgggat ctgccattgt cagacatgta taccccgtac gtcttcccga gcgaaaacgg
     5821 tctgcgctgc gggacgcgcg aattgaatta tggcccacac cagtggcgcg gcgacttcca
     5881 gttcaacatc agccgctaca gtcaacagca actgatggaa accagccatc gccatctgct
     5941 gcacgcggaa gaaggcacat ggctgaatat cgacggtttc catatgggga ttggtggcga
     6001 cgactcctgg agcccgtcag tatcggcgga attccagctg agcgccggtc gctaccatta
     6061 ccagttggtc tggtgtcaaa aataataata accgggcagg ggggatccgc agatcccggc
     6121 cagataccga tgctgccgca gcaaaagcag gagcagatgc cgccgtcgca ggcgaagatg
     6181 tcgcagacgg aggaggcgat gctgccggcg gaggaggcga agtaagtaga gggctgggct
     6241 gggctgtggg gggtgtgggg tgcgggactg ggcagtctgg gagtccctct caccactttt
     6301 cttacctttc taggatgctg ccgtcgccgc cgctcataca ccataaggtg taaaaaatac
     6361 tagatgcaca gaatagcaag tccatcaaaa ctcctgcgtg agaattttac cagacttcaa
     6421 gagcatctcg ccacatcttg aaaaatgcca ccgtccgatg aaaaacagga gcctgctaag
     6481 gaacaatgcc acctgtcaat aaatgttgaa aactcatccc attcctgcct cttggtcctt
     6541 gggcttgggg aggggtgcgc ggatgtggtt agggaacatg actggtcaaa tgggaagggc
     6601 ttcaaaagaa ttcccaatat tgactaccaa gccacctgta cagatctgcc tgcaggcatg
     6661 caagcttggc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     6721 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     6781 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt
     6841 attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa
     6901 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     6961 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     7021 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cg
//
