LOCUS       sequence-182530-        7242 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of Glt1b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7242)
  AUTHORS   Chen W, Aoki C, Mahadomrongkul V, Gruber CE, Wang GJ, Blitzblau R, 
            Irwin N, Rosenberg PA
  TITLE     Expression of a variant form of the glutamate transporter GLT1 in 
            neuronal cultures and in neurons and astrocytes in the rat brain.
  JOURNAL   J Neurosci. 2002 Mar 15;22(6):2142-52.
  PUBMED    11896154
REFERENCE   2  (bases 1 to 7242)
  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."; date: "2002-03-15"; volume: "22(6)"; pages: "2142-52"
COMMENT     Sequence Label: pcDNA3 Glt1b
FEATURES             Location/Qualifiers
     source          1..7242
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             22..816
                     /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(76..95)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     686..705
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    990..1111
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1027..1046)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1081..1100
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1160..1176)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1160..1176)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1173..1195)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1184..1200
                     /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(1208..1214,1215..1232,1233..1238))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1253..1274
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1391..1408)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1562..2150)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1642..1661)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2321..3112,3113..3181))
                     /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     2944..2963
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3182..3286)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3361..3380)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3552..3931
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3932..4135
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4086..4106
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4180..4199
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4180..4198
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(6112..6130)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6113..6130)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(6150..6167)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6156..6380
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6426..6854
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6513..6532)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6723..6744
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6863..6883)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6868..7197
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7048..7183
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7110..7129
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
       61 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
      121 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
      181 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
      241 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
      301 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
      361 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
      421 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
      481 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
      541 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
      601 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
      661 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
      721 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
      781 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
      841 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
      901 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
      961 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     1021 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     1081 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     1141 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     1201 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     1261 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     1321 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     1381 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     1441 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     1501 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     1561 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     1621 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     1681 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     1741 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     1801 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     1861 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     1921 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     1981 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     2041 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     2101 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     2161 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     2221 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     2281 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     2341 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     2401 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     2461 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     2521 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     2581 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     2641 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     2701 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     2761 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     2821 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     2881 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     2941 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     3001 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     3061 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     3121 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     3181 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     3241 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     3301 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag
     3361 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
     3421 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
     3481 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag
     3541 atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
     3601 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     3661 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     3721 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     3781 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     3841 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     3901 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     3961 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     4021 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     4081 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctctg
     4141 gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc actataggga
     4201 gacccaagct tggtaccgag ctcggatcca ctagtaacgg ccgccagtgt gctggaattc
     4261 gcccttcgcc atggcatcaa ccgagggtgc caacaatatg cccaagcagg tggaagtgcg
     4321 catgcacgac agccacctca gctccgagga gccaaagcac cgaaacctgg gcatgcgcat
     4381 gtgcgacaag ctggggaaga acctcctgct ctcactgact gtgtttggtg tcatcctggg
     4441 agcagtatgt ggcgggctgc ttcgcttggc ggctcccatc caccctgatg tggtcatgtt
     4501 gatagccttc ccgggggaca tcctcatgag gatgctgaag atgctcatcc tccctctcat
     4561 catttccagt ctcatcacag ggctgtcagg gctggatgct aaagccagcg gccgcctagg
     4621 cacgagagcc atggtatatt acatgtccac gaccatcatt gccgctgtgc tgggggtcat
     4681 cctggtgttg gccatccacc ctgggaatcc caaactcaag aagcagctgg ggcctgggaa
     4741 gaagaacgac gaggtgtcca gcctggatgc cttcctggat ctcattagaa atctcttccc
     4801 ggagaacctg gtacaagcct gtttccaaca gattcagact gtgacaaaga aagttctggt
     4861 ggcacctcca tccgaggagg ccaatacaac caaggcagtc atctccctgt tgaatgagac
     4921 catgaatgag gcccctgaag aaactaagat cgttatcaag aagggcctgg agttcaagga
     4981 cgggatgaat gtcttaggtc tgattggatt ctttattgct ttcggcattg ccatggggaa
     5041 gatgggtgag caggccaagc tgatggtgga gttcttcaac attctgaacg agattgtcat
     5101 gaagttagtg atcatgatca tgtggtattc cccgctgggt atcgcctgct tgatctgtgg
     5161 gaagatcatc gccatcaagg acttagaagt ggttgctagg cagctgggga tgtacatgat
     5221 cacagtgatc gtgggcctca tcattcatgg gggcatcttt ctccccttga tttactttgt
     5281 agtgaccaga aaaaacccat tctccttttt tgctggcatt ttccaagcct ggatcactgc
     5341 cctgggaacc gcttccagtg ctggaacttt gcctgtcacc ttccgttgct tggaagataa
     5401 tctagggatt gacaagcgtg tgaccagatt cgtcctccca gtcggagcaa ccattaacat
     5461 ggatggtaca gccctttacg aagccgtggc agccatcttc atagcccaga tgaatggggt
     5521 catcctggat ggaggtcaga tagtgactgt aagccttaca gcaactctgg cgagcattgg
     5581 tgcagccagt attcccagcg ccgggttggt caccatgctc ctcattctca cagctgtggg
     5641 cctgccgaca gaggacatca gtctgctggt ggcagtggac tggctgctgg atagaatgag
     5701 aacttcggtc aatgtagtgg gcgattcttt tggggctggg attgtctatc acctttccaa
     5761 gtccgagctg gacaccattg actcccaaca ccgaatgcac gaagacatcg aaatgaccaa
     5821 gacgcagtcc gtttatgacg acacgaagaa ccacagggaa agcaactcta atcagtgtgt
     5881 ctatgccgca cacaactctg tcgtaataga tgagtgcaag gtacctttcc cattcctgga
     5941 tatcgagacc tgcatatgaa tagtgcatgc tggactctta caaaaaagca ccatcccctg
     6001 catccattct taccattctt tgtaaggaat catgtcaccc aagctaaggg cgaattctgc
     6061 agatatccat cacactggcg gccgctcgag catgcatcta gagggcccta ttctatagtg
     6121 tcacctaaat gctagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     6181 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     6241 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     6301 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     6361 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     6421 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     6481 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     6541 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     6601 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     6661 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     6721 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     6781 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     6841 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     6901 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     6961 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     7021 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     7081 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     7141 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     7201 ctcccgggag cttgtatatc cattttcgga tctgatcaag ag
//
