LOCUS       sequence-234650-        6113 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GABAA receptor expression (gamma 1 subunit with EGFP tag at the 
            C-terminus).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6113)
  AUTHORS   de Blas, Angel
  TITLE     GABAA receptor subunit plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6113)
  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            "Unpublished"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: gamma1-EGFP
FEATURES             Location/Qualifiers
     source          1..6113
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2053..2062
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2059..2061,2062..2064,2065..2778)
                     /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(2059..2061,2062..2064,2065..2775)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2104..2125)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2365..2384)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2712..2733
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2901..3022
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2938..2957)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2992..3011
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3029..3484)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3166..3187)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3378..3397
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3511..3615
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3617..3974
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3617..3808
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3640..3660)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3825..3960
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4009..4803
                     /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             4009..4803
                     /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(4063..4082)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4673..4692
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4991..5010)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5035..5082
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5411..5999
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5900..5919
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttatg ggttctggga aagtcttcct tttctctcct
      661 tccctcctgt ggagtcaaac tagaggagtg aggttgatat tcttgttact aaccctgcat
      721 ctgggaaact gcattgataa agcagatgat gaagatgatg aagatttaac tatgaacaaa
      781 acatgggtct tggcacctaa aattcatgaa ggagatatca cacagattct caactcttta
      841 cttcaaggct atgataacaa acttcgtcca gatataggcg tgagacccac agtaattgaa
      901 actgatgttt atgtaaacag cattggacct gttgatccca taaacatgga gtacacgata
      961 gacatcattt ttgcccagac ctggtttgat agccgtttaa agttcaacag caccatgaaa
     1021 gtccttatgc ttaacagcaa catggtggga aaaatttgga tccctgacac gttcttcagg
     1081 aactcaagga aatctgatgc gcactggata acaacgccca atcgcctgct gcggatatgg
     1141 agtgatggaa gggttctcta cactctgaga ttaacaatta atgcagaatg ctaccttcaa
     1201 ctccataact ttcctatgga tgaacattcc tgtccactgg aattttcaag ctatggatac
     1261 cctaaaaatg aaattgagta caagtggaaa aagccctcag tggaagtagc tgatcctaag
     1321 tactggagat tgtatcagtt tgcctttgta gggttaagaa attcaaccga aatctctcac
     1381 acaatctccg gagattatat tatcatgact atcttttttg acctgagcag acggatgggc
     1441 tatttcacaa ttcagaccta cattccgtgc attctaacag ttgttctttc ctgggtgtct
     1501 ttttggatca acaaagatgc tgtacctgct agaacatccc tgggtatcac tacggttttg
     1561 actatgacaa ccctcagtac aatcgctaga aaatctttac ctaaggtttc ttacgtgaca
     1621 gcaatggatc tctttgtctc tgtgtgtttc atttttgtgt ttgcagcact catggagtat
     1681 ggaaccttgc attattttac tagtaacaat aaaggaaaaa ccaccagaga caggaagctg
     1741 aaaagcaaaa cttcggtatc cccaggtctc catgctggat ctactctgat tcccatgaac
     1801 aacatttcta tgcctcaagg ggaagatgat tatggttacc agtgcttgga gggcaaagac
     1861 tgtgctacct ttttctgttg ctttgaagac tgcagaactg ggtcctggag agaagggcga
     1921 atacacatac gcattgccaa aatcgactcc tactccagga tctttttccc aacagctttt
     1981 gccttgttca atctggttta ctgggttgga tatctatact taacggtacc gcgggcccgg
     2041 gatccaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     2101 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     2161 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     2221 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
     2281 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     2341 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     2401 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     2461 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
     2521 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
     2581 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     2641 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag
     2701 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     2761 gacgagctgt acaagtaaag cggccgcgac tctagatcat aatcagccat accacatttg
     2821 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     2881 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     2941 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     3001 ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc
     3061 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     3121 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     3181 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     3241 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     3301 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     3361 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     3421 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     3481 gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     3541 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     3601 tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt
     3661 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3721 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3781 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     3841 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     3901 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     3961 taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     4021 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     4081 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     4141 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca
     4201 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     4261 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     4321 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     4381 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     4441 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     4501 ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc
     4561 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     4621 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     4681 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     4741 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     4801 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     4861 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     4921 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg
     4981 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     5041 taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt
     5101 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
     5161 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
     5221 acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt tagattgatt
     5281 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5341 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     5401 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     5461 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     5521 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     5581 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     5641 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     5701 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     5761 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     5821 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     5881 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     5941 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6001 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     6061 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg cat
//
