LOCUS       sequence-228581-        6190 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GABAA receptor expression (alpha1 subunit). EGFP tag at the 
            C-terminus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6190)
  AUTHORS   Christie SB, Li RW, Miralles CP, Yang B-, De Blas AL
  TITLE     Clustered and non-clustered GABAA receptors in cultured hippocampal 
            neurons.
  JOURNAL   Mol Cell Neurosci. 2006 Jan;31(1):1-14. doi: 
            10.1016/j.mcn.2005.08.014. Epub 2005 Sep 21.
  PUBMED    16181787
REFERENCE   2  (bases 1 to 6190)
  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: 
            "10.1016/j.mcn.2005.08.014"; journalName: "Mol Cell Neurosci"; date:
            "2006-01"; volume: "31"; issue: "1"; pages: "1-14"
COMMENT     Sequence Label: alpha1-EGFP
FEATURES             Location/Qualifiers
     source          1..6190
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        119..422
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        423..626
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     577..597
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2188..2197
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2194..2196,2197..2199,2200..2913)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2194..2196,2197..2199,2200..2910)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2239..2260)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2500..2519)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2847..2868
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3036..3157
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3073..3092)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3127..3146
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3164..3619)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3301..3322)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3513..3532
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3646..3750
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3752..4109
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3752..3943
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3775..3795)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3960..4095
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4144..4938
                     /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             4144..4938
                     /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(4198..4217)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4808..4827
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5126..5145)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5170..5217
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5546..6134
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6035..6054
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta
       61 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      121 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      181 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      241 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      301 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      361 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      421 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      481 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      541 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      601 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc
      661 ggactcagat ctcgagctca agcttctaga gatccctcga cctcgagatc cattgtgctc
      721 tgaattcgtc gagacgcctc ccgggtctgg agcgatccgg tgcccagagg gggccccgag
      781 ctggacaagc ccgtgatgaa gaaaagtcgg ggtctctctg actatctttg ggcctggacc
      841 ctcattctga gcactctctc gggaagaagc tatggacagc cctcccaaga tgaacttaag
      901 gacaacacca ctgtcttcac gaggattttg gaccgactgc tggatggtta tgacaatcgt
      961 ctgagaccag gcttgggaga gcgtgtaact gaagtgaaga cggacatctt tgtcaccagt
     1021 ttcggacccg tgtcagacca cgatatggaa tatacaatag atgtgttttt ccgccaaagc
     1081 tggaaggatg aaagattaaa attcaaagga cccatgacag tgctccggct gaacaacctg
     1141 atggccagta aaatctggac tccagataca tttttccaca atggaaaaaa gtctgtggcc
     1201 cacaacatga ccatgcccaa taaactcctg cgtatcacag aggatggcac actgctgtac
     1261 accatgaggt tgactgtgag agccgaatgc cccatgcact tagaagactt tcccatggat
     1321 gcccatgcct gcccactaaa atttgggagc tatgcttata caagagcaga agttgtctat
     1381 gagtggacaa gggagccagc ccgctcagtg gttgtggcag aagatgggtc acgtttaaac
     1441 cagtatgacc ttcttgggca aacagttgac tctggaattg ttcagtccag tactggagaa
     1501 tatgtggtta tgacgactca ctttcacttg aagagaaaaa tcggctactt tgttattcaa
     1561 acatatctgc catgcataat gacagtcatt ctctcccaag tctccttctg gcttaacaga
     1621 gagtcagtac cagcaagaac tgtctttgga gtgacgaccg ttctgaccat gacaaccttg
     1681 agtatcagtg ccagaaattc cctcccaaag gtggcttatg caacggccat ggactggttt
     1741 attgcagtgt gctatgcctt cgtgttctcg gctctgattg agtttgccac agtaaactat
     1801 ttcaccaaga gagggtatgc gtgggatggc aaaagcgtgg ttccagaaaa gccaaagaaa
     1861 gtgaaggatc ctctcattaa gaaaaacaac acatatgctc ctacagcaac cagctatacc
     1921 cctaacttag ccaggggtga ccccggcttg gcaactattg ctaaaagtgc gaccatagaa
     1981 ccgaaagaag tcaagcctga gacaaaaccg ccagaaccca agaaaacctt taacagcgtc
     2041 agcaaaatcg accgactgtc aagaatagcc tttccgctgc tatttggaat ctttaactta
     2101 gtctattggg ccacgtattt aaacagagag cctcagctaa aagcccccac accccatcag
     2161 gtaccgcggg cccgggatcc accggtcgcc accatggtga gcaagggcga ggagctgttc
     2221 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     2281 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     2341 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     2401 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     2461 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     2521 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     2581 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     2641 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     2701 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     2761 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     2821 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     2881 atcactctcg gcatggacga gctgtacaag taaagcggcc gcgactctag atcataatca
     2941 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     3001 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     3061 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     3121 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat
     3181 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     3241 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     3301 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     3361 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     3421 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     3481 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     3541 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     3601 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3661 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3721 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     3781 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3841 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3901 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3961 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4021 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     4081 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     4141 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     4201 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     4261 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     4321 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     4381 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     4441 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     4501 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     4561 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     4621 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     4681 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     4741 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     4801 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     4861 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4921 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     4981 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     5041 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     5101 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     5161 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     5221 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     5281 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     5341 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     5401 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     5461 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     5521 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     5581 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     5641 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     5701 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     5761 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     5821 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     5881 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     5941 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     6001 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     6061 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     6121 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     6181 cttttgctca
//
