LOCUS       sequence-228584-        6110 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GABAA receptor expression (alpha5 subunit). EGFP tag at the 
            C-terminus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6110)
  TITLE     GABAA receptor expression
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6110)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: alpha5-EGFP
FEATURES             Location/Qualifiers
     source          1..6110
                     /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      2108..2117
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2114..2116,2117..2119,2120..2833)
                     /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(2114..2116,2117..2119,2120..2830)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2159..2180)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2420..2439)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2767..2788
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2956..3077
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2993..3012)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3047..3066
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3084..3539)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3221..3242)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3433..3452
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3566..3670
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3672..4029
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3672..3863
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3695..3715)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3880..4015
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4064..4858
                     /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             4064..4858
                     /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(4118..4137)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4728..4747
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5046..5065)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5090..5137
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5466..6054
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5955..5974
                     /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 agcttatgga caatggaatg ctctctagat ttatcatgac
      721 caaaaccctc cttgtcttct gcatttccat gaccttatcc agtcactttg gcttttcaca
      781 aatgccaact agttctgtac aagatgagac caatgacaac atcacaatat tcaccaggat
      841 cttggacggg ctcttggatg gctatgacaa cagactgcgg cctgggctgg gagagcgaat
      901 cacgcaggtg cgaacagaca tctatgttac cagctttggc ccagtgtccg acacggaaat
      961 ggaatatacc atagatgtgt ttttccgaca aagctggaaa gatgaaaggc tgcggtttaa
     1021 ggggcctatg caacgtctcc ctctcaacaa ccttcttgcc agcaaaatct ggaccccaga
     1081 cacattcttc cacaatggga agaagtccat tgcgcacaac atgacgacac ccaacaagct
     1141 gctgaggctg gaggatgatg gcacacttct ctacaccatg cgcctgacga tctctgctga
     1201 gtgtccaatg cagcttgagg acttcccgat ggatgcccat gcttgtcccc tgaaatttgg
     1261 cagttatgct taccctaatt cggaagttgt ctatgtttgg accaatggtt ccaccaagtc
     1321 tgtggtggtg gcagaagatg gctccagact caaccagtac cacctcatgg ggcagacagt
     1381 aggcactgag aacatcagca ccagcacagg tgaatataca atcatgactg ctcattttca
     1441 cctgaagagg aagatcgggt actttgtcat ccagacgtac cttccctgca tcatgactgt
     1501 catcttatcc caggtgtctt tttggcttaa tcgagaatct gtcccagcta ggacagtttt
     1561 tggagtgacc acagtgctga ccatgacaac cctcagcatc agtgcccgga attcgctgcc
     1621 caaagtggcc tatgccacag ccatggactg gttcattgct gtctgctatg catttgtctt
     1681 ctctgccctg attgaatttg ccacagtcaa ctactttaca aagagaggat gggcctggga
     1741 tggcaagaag gccttggaag cagctaaaat caagaaaaaa gaacgtgaac tcatactaaa
     1801 taagtcaaca aatgctttta caactgggaa gttgacccat cctccaaaca tcccaaagga
     1861 gcagcttcca ggcgggactg ggaatgctgt gggtacagcc tcaatcagag catctgagga
     1921 gaagacttct gagagtaaaa agacctacaa cagcatcagc aagatcgaca aaatgtcccg
     1981 gattgtgttc cccattttgt ttggcacttt caatctagtt tactgggcaa catatttgaa
     2041 tagggagccc gtgataaaag gggctacctc tccaaagtcg gtaccgcggg cccgggatcc
     2101 accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     2161 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     2221 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     2281 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
     2341 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     2401 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     2461 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     2521 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     2581 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     2641 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     2701 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     2761 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     2821 gctgtacaag taaagcggcc gcgactctag atcataatca gccataccac atttgtagag
     2881 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     2941 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     3001 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     3061 ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt
     3121 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     3181 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     3241 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     3301 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     3361 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     3421 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     3481 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     3541 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     3601 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3661 aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     3721 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3781 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3841 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3901 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3961 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     4021 ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     4081 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     4141 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     4201 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg
     4261 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     4321 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     4381 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     4441 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     4501 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     4561 gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt
     4621 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     4681 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     4741 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     4801 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     4861 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     4921 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     4981 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg
     5041 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     5101 agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     5161 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     5221 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     5281 ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa
     5341 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     5401 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5461 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5521 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     5581 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     5641 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     5701 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     5761 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     5821 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     5881 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     5941 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6001 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     6061 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
//
