LOCUS       sequence-228639-        6121 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GABAA receptor expression (gamma2 short subunit) double tagged, near
            N terminus and at C-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6121)
  AUTHORS   de Blas, Angel
  TITLE     GABAA receptor subunit plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6121)
  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            "Unpublished"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: Myc-gamma2s-EGFP
FEATURES             Location/Qualifiers
     source          1..6121
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             663..692
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      2001..2010
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2007..2009,2010..2012,2013..2726)
                     /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(2007..2009,2010..2012,2013..2723)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2052..2073)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2313..2332)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2660..2681
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2849..2970
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2886..2905)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2940..2959
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2977..3432)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3114..3135)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3326..3345
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3459..3563
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3565..3922
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3565..3756
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3588..3608)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3773..3908
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3957..4751
                     /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             3957..4751
                     /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(4011..4030)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4621..4640
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4939..4958)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4983..5030
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5359..5947
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5848..5867
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcatga
      541 gttcgccaaa tacatggagc actggaagca cagtctactc tcctgtattt tcacagaaaa
      601 tgacgctgtg gattctgctc ctgctatcgc tctacccagg cttcactagc caaaagtcag
      661 atgaacaaaa actaatatca gaagaagacc tagatgacta tgaagattat gcttctaata
      721 aaacatgggt gttgactcca aaagttcccg agggtgatgt cactgtcatc ttaaacaacc
      781 ttctggaagg gtacgacaac aaacttcggc ccgacatagg agtgaaacca acattaattc
      841 atacagatat gtacgtgaac agcattggtc cagtgaatgc tatcaatatg gaatacacaa
      901 ttgatatttt ttttgcccaa acctggtatg acagacgttt gaaatttaac agtaccatta
      961 aagttctccg attgaatagc aatatggtgg ggaaaatctg gattccagac actttcttca
     1021 ggaactccaa aaaagcggat gctcactgga tcacgactcc caacaggatg ctgagaattt
     1081 ggaatgacgg tcgagttctc tacaccttaa ggctaacaat tgatgccgag tgccagttgc
     1141 aattacacaa cttcccaatg gatgaacact cctgccccct ggagttctcc agttatggtt
     1201 atcctcgtga agaaattgtt tatcaatgga agcgcagttc tgttgaagtg ggagacacaa
     1261 ggtcatggag gctgtatcag ttttcctttg ttggattgag gaataccact gaagtagtga
     1321 agacaacttc tggtgactat gtggttatgt ccgtgtactt tgatctgagc agaagaatgg
     1381 ggtactttac catccagacc tacattccct gcacactcat tgtggttctg tcctgggtat
     1441 ccttctggat caataaggat gctgtccctg caagaacatc tttaggaatc acgactgtcc
     1501 tgacgatgac cactctcagc accatagccc ggaagtctct gcccaaggtc tcctatgtca
     1561 cagcaatgga tctcttcgtc tctgtttgct tcatctttgt gttttcagct ttggtggagt
     1621 atggtaccct gcactatttt gtgagcaacc ggaaaccaag caaggataaa gacaaaaaga
     1681 agaaaaaccc tgcccctacc attgatatcc gtcccagatc agcaacgatc caaatgaaca
     1741 atgccaccca ccttcaagag agggatgaag aatatggcta tgagtgtttg gatggcaagg
     1801 actgtgccag tttcttttgc tgttttgaag actgccgaac aggagcctgg agacacggga
     1861 ggatacacat tcgcattgcc aaaatggact cctatgctcg gatcttcttc cctaccgcct
     1921 tctgcttgtt caatcttgtt tactgggtct cctatcttta tctgcagtcg acggtaccgc
     1981 gggcccggga tccaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg
     2041 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     2101 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     2161 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     2221 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     2281 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     2341 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     2401 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     2461 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     2521 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     2581 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
     2641 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     2701 tcggcatgga cgagctgtac aagtaaagcg gccgcgactc tagatcataa tcagccatac
     2761 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     2821 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     2881 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     2941 tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt
     3001 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     3061 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     3121 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     3181 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     3241 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     3301 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     3361 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     3421 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     3481 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3541 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     3601 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3661 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3721 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     3781 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     3841 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     3901 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
     3961 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     4021 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     4081 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     4141 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     4201 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     4261 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     4321 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     4381 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     4441 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     4501 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     4561 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     4621 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     4681 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     4741 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     4801 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     4861 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     4921 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     4981 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
     5041 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     5101 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     5161 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
     5221 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     5281 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     5341 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5401 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5461 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     5521 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     5581 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     5641 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     5701 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     5761 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     5821 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     5881 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     5941 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     6001 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca
     6061 ttagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
     6121 g
//
