LOCUS       sequence-322412-        6437 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Rat GLT-1a cDNA N-terminally tagged with EGFP with a v5 tag in the 
            extracellular loop.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6437)
  AUTHORS   Al Awabdh S, Gupta-Agarwal S, Sheehan DF, Muir J, Norkett R, 
            Twelvetrees AE, Griffin LD, Kittler JT
  TITLE     Neuronal activity mediated regulation of glutamate transporter GLT-1
            surface diffusion in rat astrocytes in dissociated and slice 
            cultures.
  JOURNAL   Glia. 2016 Jul;64(7):1252-64. doi: 10.1002/glia.22997.
  PUBMED    27189737
REFERENCE   2  (bases 1 to 6437)
  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"; doi: "10.1002/glia"; 
            journalName: "Glia"; date: "2016-07"; volume: "64"; issue: "7"; 
            pages: "1252-64"
COMMENT     Sequence Label: GFP-GLT1b-V5
FEATURES             Location/Qualifiers
     source          1..6437
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        53..356
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        357..560
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     511..531
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      599..608
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(605..607,608..610,611..1321)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(605..607,608..610,611..1321)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(650..671)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(911..930)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1258..1279
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1955..1996
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    3217..3338
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3254..3273)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3308..3327
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3345..3800)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3482..3503)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3694..3713
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3827..3931
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3933..4290
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3933..4124
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3956..3976)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4141..4276
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4325..5119
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4325..5119
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4379..4398)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4989..5008
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5307..5326)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5351..5398
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5727..6315
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6216..6235
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
       61 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      121 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      181 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      241 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      301 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
      361 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
      421 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
      481 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
      541 aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgc taccggtcgc
      601 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
      661 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
      721 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
      781 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
      841 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
      901 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
      961 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     1021 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     1081 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     1141 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1201 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     1261 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     1321 gtccggactc agatctcgag ctcaagcttc gaattctatg gcatcaaccg agggtgccaa
     1381 caatatgccc aagcaggtgg aagtgcgcat gcacgacagc cacctcagct ccgaggagcc
     1441 aaagcaccga aacctgggca tgcgcatgtg cgacaagctg gggaagaacc tcctgctctc
     1501 actgactgtg tttggtgtca tcctgggagc agtatgtggc gggctgcttc gcttggcggc
     1561 tcccatccac cctgatgtgg tcatgttgat agccttcccg ggggacatcc tcatgaggat
     1621 gctgaagatg ctcatcctcc ctctcatcat ttccagtctc atcacagggc tgtcagggct
     1681 ggatgctaaa gccagcggcc gcctaggcac gagagccatg gtatattaca tgtccacgac
     1741 catcattgcc gctgtgctgg gggtcatcct ggtgttggcc atccaccctg ggaatcccaa
     1801 actcaagaag cagctggggc ctgggaagaa gaacgacgag gtgtccagcc tggatgcctt
     1861 cctggatctc attagaaatc tcttcccgga gaacctggta caagcctgtt tccaacagat
     1921 tcagactgtg acaaagaaag ttctggtggc acctggcaag cccatcccca accccctgct
     1981 gggcctggac agcaccccat ccgaggaggc caatacaacc aaggcagtca tctccctgtt
     2041 gaatgagacc atgaatgagg cccctgaaga aactaagatc gttatcaaga agggcctgga
     2101 gttcaaggac gggatgaatg tcttaggtct gattggattc tttattgctt tcggcattgc
     2161 catggggaag atgggtgagc aggccaagct gatggtggag ttcttcaaca ttctgaacga
     2221 gattgtcatg aagttagtga tcatgatcat gtggtattcc ccgctgggta tcgcctgctt
     2281 gatctgtggg aagatcatcg ccatcaagga cttagaagtg gttgctaggc agctggggat
     2341 gtacatgatc acagtgatcg tgggcctcat cattcatggg ggcatctttc tccccttgat
     2401 ttactttgta gtgaccagaa aaaacccatt ctcctttttt gctggcattt tccaagcctg
     2461 gatcactgcc ctgggaaccg cttccagtgc tggaactttg cctgtcacct tccgttgctt
     2521 ggaagataat ctagggattg acaagcgtgt gaccagattc gtcctcccag tcggagcaac
     2581 cattaacatg gatggtacag ccctttacga agccgtggca gccatcttca tagcccagat
     2641 gaatggggtc atcctggatg gaggtcagat agtgactgta agccttacag caactctggc
     2701 gagcattggt gcagccagta ttcccagcgc cgggttggtc accatgctcc tcattctcac
     2761 agctgtgggc ctgccgacag aggacatcag tctgctggtg gcagtggact ggctgctgga
     2821 tagaatgaga acttcggtca atgtagtggg cgattctttt ggggctggga ttgtctatca
     2881 cctttccaag tccgagctgg acaccattga ctcccaacac cgaatgcacg aagacatcga
     2941 aatgaccaag acgcagtccg tttatgacga cacgaagaac cacagggaaa gcaactctaa
     3001 tcagtgtgtc tatgccgcac acaactctgt cgaatagatg agtgcaaggt acctttccca
     3061 ttcctggata tcgagacctg catatgagga tccaccggat ctagataact gatcataatc
     3121 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     3181 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     3241 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     3301 tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtaaattg taagcgttaa
     3361 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     3421 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     3481 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     3541 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     3601 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     3661 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     3721 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     3781 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     3841 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     3901 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
     3961 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4021 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     4081 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     4141 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4201 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     4261 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt
     4321 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     4381 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     4441 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     4501 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     4561 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     4621 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     4681 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     4741 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     4801 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
     4861 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     4921 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     4981 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     5041 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     5101 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     5161 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     5221 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     5281 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     5341 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
     5401 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     5461 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     5521 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
     5581 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     5641 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5701 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5761 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     5821 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     5881 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     5941 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     6001 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     6061 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     6121 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     6181 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     6241 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     6301 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     6361 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     6421 gccatgcatt agttatt
//
