LOCUS       sequence-321937-        6429 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Rat GLT-1a cDNA N-terminally tagged with EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6429)
  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 6429)
  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
FEATURES             Location/Qualifiers
     source          1..6429
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        72..375
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        376..579
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     530..550
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      618..627
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(624..626,627..629,630..1340)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(624..626,627..629,630..1340)
                     /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(669..690)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(930..949)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1277..1298
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3228..3349
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3265..3284)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3319..3338
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3356..3811)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3493..3514)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3705..3724
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3838..3942
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3944..4301
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3944..4135
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3967..3987)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4152..4287
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4336..5130
                     /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             4336..5130
                     /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(4390..4409)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5000..5019
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5318..5337)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5362..5409
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5738..6326
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6227..6246
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccgccatgca ttagttatta atagtaatca attacggggt cattagttca tagcccatat
       61 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      121 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      181 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      241 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      301 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      361 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      421 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      481 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      541 ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc
      601 cgctagcgct accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg
      661 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
      721 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
      781 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
      841 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
      901 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
      961 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     1021 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     1081 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     1141 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     1201 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
     1261 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     1321 gcatggacga gctgtacaag tccggactca gatctcgagc tcaagcttcg aattctatgg
     1381 catcaaccga gggtgccaac aatatgccca agcaggtgga agtgcgcatg cacgacagcc
     1441 acctcagctc cgaggagcca aagcaccgaa acctgggcat gcgcatgtgc gacaagctgg
     1501 ggaagaacct cctgctctca ctgactgtgt ttggtgtcat cctgggagca gtatgtggcg
     1561 ggctgcttcg cttggcggct cccatccacc ctgatgtggt catgttgata gccttcccgg
     1621 gggacatcct catgaggatg ctgaagatgc tcatcctccc tctcatcatt tccagtctca
     1681 tcacagggct gtcagggctg gatgctaaag ccagcggccg cctaggcacg agagccatgg
     1741 tatattacat gtccacgacc atcattgccg ctgtgctggg ggtcatcctg gtgttggcca
     1801 tccaccctgg gaatcccaaa ctcaagaagc agctggggcc tgggaagaag aacgacgagg
     1861 tgtccagcct ggatgccttc ctggatctca ttagaaatct cttcccggag aacctggtac
     1921 aagcctgttt ccaacagatt cagactgtga caaagaaagt tctggtggca cctccatccg
     1981 aggaggccaa tacaaccaag gcagtcatct ccctgttgaa tgagaccatg aatgaggccc
     2041 ctgaagaaac taagatcgtt atcaagaagg gcctggagtt caaggacggg atgaatgtct
     2101 taggtctgat tggattcttt attgctttcg gcattgccat ggggaagatg ggtgagcagg
     2161 ccaagctgat ggtggagttc ttcaacattc tgaacgagat tgtcatgaag ttagtgatca
     2221 tgatcatgtg gtattccccg ctgggtatcg cctgcttgat ctgtgggaag atcatcgcca
     2281 tcaaggactt agaagtggtt gctaggcagc tggggatgta catgatcaca gtgatcgtgg
     2341 gcctcatcat tcatgggggc atctttctcc ccttgattta ctttgtagtg accagaaaaa
     2401 acccattctc cttttttgct ggcattttcc aagcctggat cactgccctg ggaaccgctt
     2461 ccagtgctgg aactttgcct gtcaccttcc gttgcttgga agataatcta gggattgaca
     2521 agcgtgtgac cagattcgtc ctcccagtcg gagcaaccat taacatggat ggtacagccc
     2581 tttacgaagc cgtggcagcc atcttcatag cccagatgaa tggggtcatc ctggatggag
     2641 gtcagatagt gactgtaagc cttacagcaa ctctggcgag cattggtgca gccagtattc
     2701 ccagcgccgg gttggtcacc atgctcctca ttctcacagc tgtgggcctg ccgacagagg
     2761 acatcagtct gctggtggca gtggactggc tgctggatag aatgagaact tcggtcaatg
     2821 tagtgggcga ttcttttggg gctgggattg tctatcacct ttccaagtcc gagctggaca
     2881 ccattgactc ccaacaccga atgcacgaag acatcgaaat gaccaagacg cagtccgttt
     2941 atgacgacac gaagaaccac agggaaagca actctaatca gtgtgtctat gccgcacaca
     3001 actctgtcgt aatagatgag tgcaaggtaa ctctggcggc caatggaaag tcagctgact
     3061 gcagtgttga ggaagaacct tggaaacgtg aaaaataagg atccaccgga tctagataac
     3121 tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     3181 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     3241 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     3301 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt
     3361 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     3421 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     3481 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     3541 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     3601 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     3661 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     3721 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     3781 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     3841 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     3901 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
     3961 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4021 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     4081 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     4141 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     4201 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     4261 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
     4321 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     4381 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     4441 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     4501 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4561 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     4621 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4681 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4741 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4801 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4861 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4921 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4981 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     5041 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     5101 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     5161 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     5221 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     5281 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac
     5341 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
     5401 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     5461 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     5521 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag
     5581 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     5641 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     5701 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     5761 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     5821 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     5881 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     5941 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     6001 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     6061 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     6121 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     6181 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     6241 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     6301 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     6361 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     6421 accgtatta
//
