LOCUS       sequence-321944-        6471 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 6471)
  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 6471)
  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-GLT1a-V5
FEATURES             Location/Qualifiers
     source          1..6471
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        109..412
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        413..616
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     567..587
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      655..664
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(661..663,664..666,667..1377)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(661..663,664..666,667..1377)
                     /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(706..727)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(967..986)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1314..1335
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2011..2052
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    3307..3428
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3344..3363)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3398..3417
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3435..3890)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3572..3593)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3784..3803
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3917..4021
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4023..4380
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4023..4214
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4046..4066)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4231..4366
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4415..5209
                     /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             4415..5209
                     /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(4469..4488)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5079..5098
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5397..5416)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5441..5488
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5817..6405
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6306..6325
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta gttattaata
       61 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
      121 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      181 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      241 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
      301 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
      361 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
      421 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
      481 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
      541 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
      601 ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc ggtcgccacc
      661 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
      721 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
      781 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
      841 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
      901 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
      961 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     1021 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     1081 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     1141 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     1201 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     1261 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     1321 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtcc
     1381 ggactcagat ctcgagctca agcttcgaat tctatggcat caaccgaggg tgccaacaat
     1441 atgcccaagc aggtggaagt gcgcatgcac gacagccacc tcagctccga ggagccaaag
     1501 caccgaaacc tgggcatgcg catgtgcgac aagctgggga agaacctcct gctctcactg
     1561 actgtgtttg gtgtcatcct gggagcagta tgtggcgggc tgcttcgctt ggcggctccc
     1621 atccaccctg atgtggtcat gttgatagcc ttcccggggg acatcctcat gaggatgctg
     1681 aagatgctca tcctccctct catcatttcc agtctcatca cagggctgtc agggctggat
     1741 gctaaagcca gcggccgcct aggcacgaga gccatggtat attacatgtc cacgaccatc
     1801 attgccgctg tgctgggggt catcctggtg ttggccatcc accctgggaa tcccaaactc
     1861 aagaagcagc tggggcctgg gaagaagaac gacgaggtgt ccagcctgga tgccttcctg
     1921 gatctcatta gaaatctctt cccggagaac ctggtacaag cctgtttcca acagattcag
     1981 actgtgacaa agaaagttct ggtggcacct ggcaagccca tccccaaccc cctgctgggc
     2041 ctggacagca ccccatccga ggaggccaat acaaccaagg cagtcatctc cctgttgaat
     2101 gagaccatga atgaggcccc tgaagaaact aagatcgtta tcaagaaggg cctggagttc
     2161 aaggacggga tgaatgtctt aggtctgatt ggattcttta ttgctttcgg cattgccatg
     2221 gggaagatgg gtgagcaggc caagctgatg gtggagttct tcaacattct gaacgagatt
     2281 gtcatgaagt tagtgatcat gatcatgtgg tattccccgc tgggtatcgc ctgcttgatc
     2341 tgtgggaaga tcatcgccat caaggactta gaagtggttg ctaggcagct ggggatgtac
     2401 atgatcacag tgatcgtggg cctcatcatt catgggggca tctttctccc cttgatttac
     2461 tttgtagtga ccagaaaaaa cccattctcc ttttttgctg gcattttcca agcctggatc
     2521 actgccctgg gaaccgcttc cagtgctgga actttgcctg tcaccttccg ttgcttggaa
     2581 gataatctag ggattgacaa gcgtgtgacc agattcgtcc tcccagtcgg agcaaccatt
     2641 aacatggatg gtacagccct ttacgaagcc gtggcagcca tcttcatagc ccagatgaat
     2701 ggggtcatcc tggatggagg tcagatagtg actgtaagcc ttacagcaac tctggcgagc
     2761 attggtgcag ccagtattcc cagcgccggg ttggtcacca tgctcctcat tctcacagct
     2821 gtgggcctgc cgacagagga catcagtctg ctggtggcag tggactggct gctggataga
     2881 atgagaactt cggtcaatgt agtgggcgat tcttttgggg ctgggattgt ctatcacctt
     2941 tccaagtccg agctggacac cattgactcc caacaccgaa tgcacgaaga catcgaaatg
     3001 accaagacgc agtccgttta tgacgacacg aagaaccaca gggaaagcaa ctctaatcag
     3061 tgtgtctatg ccgcacacaa ctctgtcgta atagatgagt gcaaggtaac tctggcggcc
     3121 aatggaaagt cagctgactg cagtgttgag gaagaacctt ggaaacgtga aaaataagga
     3181 tccaccggat ctagataact gatcataatc agccatacca catttgtaga ggttttactt
     3241 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     3301 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     3361 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     3421 gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     3481 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     3541 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     3601 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     3661 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     3721 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     3781 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     3841 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
     3901 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     3961 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     4021 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     4081 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4141 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4201 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4261 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     4321 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4381 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     4441 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     4501 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     4561 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
     4621 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     4681 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     4741 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     4801 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     4861 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     4921 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     4981 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     5041 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     5101 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     5161 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     5221 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     5281 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     5341 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa
     5401 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     5461 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     5521 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     5581 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     5641 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt
     5701 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5761 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5821 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5881 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5941 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     6001 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6061 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6121 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6181 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6241 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6301 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6361 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6421 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt t
//
