LOCUS       sequence-321942-        6395 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Rat GLT-1b cDNA N-terminally tagged with EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6395)
  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 6395)
  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: "2016-07"; volume: "64"; issue: "7"; 
            pages: "1252-64"
COMMENT     Sequence Label: GFP-GLT1b
FEATURES             Location/Qualifiers
     source          1..6395
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        89..392
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        393..596
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     547..567
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      635..644
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(641..643,644..646,647..1357)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(641..643,644..646,647..1357)
                     /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(686..707)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(947..966)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1294..1315
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3211..3332
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3248..3267)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3302..3321
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3339..3794)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3476..3497)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3688..3707
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3821..3925
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3927..4284
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3927..4118
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3950..3970)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4135..4270
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4319..5113
                     /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             4319..5113
                     /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(4373..4392)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4983..5002
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5301..5320)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5345..5392
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5721..6309
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6210..6229
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgtggataac cgtattaccg ccatgcatta gttattaata gtaatcaatt acggggtcat
       61 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
      121 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
      181 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
      241 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
      301 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
      361 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg
      421 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
      481 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
      541 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt
      601 tagtgaaccg tcagatccgc tagcgctacc ggtcgccacc atggtgagca agggcgagga
      661 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
      721 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
      781 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
      841 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
      901 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
      961 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     1021 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     1081 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     1141 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     1201 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
     1261 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     1321 cgccgggatc actctcggca tggacgagct gtacaagtcc ggactcagat ctcgagctca
     1381 agcttcgaat tctatggcat caaccgaggg tgccaacaat atgcccaagc aggtggaagt
     1441 gcgcatgcac gacagccacc tcagctccga ggagccaaag caccgaaacc tgggcatgcg
     1501 catgtgcgac aagctgggga agaacctcct gctctcactg actgtgtttg gtgtcatcct
     1561 gggagcagta tgtggcgggc tgcttcgctt ggcggctccc atccaccctg atgtggtcat
     1621 gttgatagcc ttcccggggg acatcctcat gaggatgctg aagatgctca tcctccctct
     1681 catcatttcc agtctcatca cagggctgtc agggctggat gctaaagcca gcggccgcct
     1741 aggcacgaga gccatggtat attacatgtc cacgaccatc attgccgctg tgctgggggt
     1801 catcctggtg ttggccatcc accctgggaa tcccaaactc aagaagcagc tggggcctgg
     1861 gaagaagaac gacgaggtgt ccagcctgga tgccttcctg gatctcatta gaaatctctt
     1921 cccggagaac ctggtacaag cctgtttcca acagattcag actgtgacaa agaaagttct
     1981 ggtggcacct ccatccgagg aggccaatac aaccaaggca gtcatctccc tgttgaatga
     2041 gaccatgaat gaggcccctg aagaaactaa gatcgttatc aagaagggcc tggagttcaa
     2101 ggacgggatg aatgtcttag gtctgattgg attctttatt gctttcggca ttgccatggg
     2161 gaagatgggt gagcaggcca agctgatggt ggagttcttc aacattctga acgagattgt
     2221 catgaagtta gtgatcatga tcatgtggta ttccccgctg ggtatcgcct gcttgatctg
     2281 tgggaagatc atcgccatca aggacttaga agtggttgct aggcagctgg ggatgtacat
     2341 gatcacagtg atcgtgggcc tcatcattca tgggggcatc tttctcccct tgatttactt
     2401 tgtagtgacc agaaaaaacc cattctcctt ttttgctggc attttccaag cctggatcac
     2461 tgccctggga accgcttcca gtgctggaac tttgcctgtc accttccgtt gcttggaaga
     2521 taatctaggg attgacaagc gtgtgaccag attcgtcctc ccagtcggag caaccattaa
     2581 catggatggt acagcccttt acgaagccgt ggcagccatc ttcatagccc agatgaatgg
     2641 ggtcatcctg gatggaggtc agatagtgac tgtaagcctt acagcaactc tggcgagcat
     2701 tggtgcagcc agtattccca gcgccgggtt ggtcaccatg ctcctcattc tcacagctgt
     2761 gggcctgccg acagaggaca tcagtctgct ggtggcagtg gactggctgc tggatagaat
     2821 gagaacttcg gtcaatgtag tgggcgattc ttttggggct gggattgtct atcacctttc
     2881 caagtccgag ctggacacca ttgactccca acaccgaatg cacgaagaca tcgaaatgac
     2941 caagacgcag tccgtttatg acgacacgaa gaaccacagg gaaagcaact ctaatcagtg
     3001 tgtctatgcc gcacacaact ctgtcgaata gatgagtgca aggtaccttt cccattcctg
     3061 gatatcgaga cctgcatatg aggatccacc ggatctagat aactgatcat aatcagccat
     3121 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     3181 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     3241 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     3301 tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg ttaatatttt
     3361 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     3421 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     3481 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     3541 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     3601 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     3661 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     3721 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     3781 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     3841 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     3901 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
     3961 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4021 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4081 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     4141 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4201 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     4261 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     4321 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     4381 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     4441 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     4501 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     4561 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     4621 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     4681 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     4741 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     4801 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     4861 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     4921 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     4981 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     5041 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     5101 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     5161 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     5221 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     5281 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     5341 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
     5401 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     5461 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     5521 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
     5581 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5641 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5701 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5761 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5821 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5881 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5941 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     6001 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     6061 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6121 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6181 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6241 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6301 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6361 gctcacatgt tctttcctgc gttatcccct gattc
//
