LOCUS       sequence-237859-        6775 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GABAA receptor expression (chimeric rat gamma 1 subunit whose large 
            intracellular loop has been replaced with the large intracellular 
            loop of the rat gamma 2 short subunit) Myc-tag near N-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6775)
  TITLE     GABAA receptor expression plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6775)
  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            "Unpublished"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-gamma1/gamma2SIL
FEATURES             Location/Qualifiers
     source          1..6775
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..447
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        448..651
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     602..622
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     696..715
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        696..714
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             851..880
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2199..2216)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2205..2429
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2475..2903
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2562..2581)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2772..2793
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2912..2932)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2917..3246
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3097..3232
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3159..3178
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3313..4107
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3367..3386)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3977..3996
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4281..4402
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4318..4337)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4372..4391
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4451..4467)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4451..4467)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4464..4486)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4475..4491
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4499..4505,4506..4523,4524..4529))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4544..4565
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4682..4699)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4853..5441)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4933..4952)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5612..6403,6404..6472))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6235..6254
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6473..6577)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6652..6671)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat
       61 acgcgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
      121 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      181 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      241 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
      301 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      361 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      421 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
      481 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
      541 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
      601 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta
      661 actagagaac ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc
      721 caagctggct agcatgggtt ctgggaaagt cttccttttc tctccttccc tcctgtggag
      781 tcaaactaga ggagtgaggt tgatattctt gttactaacc ctgcatctgg gaaactgcat
      841 tgataaagca gaacagaagt tgatttccga agaagacctc gatgatgaag atgatgaaga
      901 tttaactatg aacaaaacat gggtcttggc acctaaaatt catgaaggag atatcacaca
      961 gattctcaac tctttacttc aaggctatga taacaaactt cgtccagata taggcgtgag
     1021 acccacagta attgaaactg atgtttatgt aaacagcatt ggacctgttg atcccataaa
     1081 catggagtac acgatagaca tcatttttgc ccagacctgg tttgatagcc gtttaaagtt
     1141 caacagcacc atgaaagtcc ttatgcttaa cagcaacatg gtgggaaaaa tttggatccc
     1201 tgacacgttc ttcaggaact caaggaaatc tgatgcgcac tggataacaa cgcccaatcg
     1261 cctgctgcgg atatggagtg atggaagggt tctctacact ctgagattaa caattaatgc
     1321 agaatgctac cttcaactcc ataactttcc tatggatgaa cattcctgtc cactggaatt
     1381 ttcaagctat ggatacccta aaaatgaaat tgagtacaag tggaaaaagc cctcagtgga
     1441 agtagctgat cctaagtact ggagattgta tcagtttgcc tttgtagggt taagaaattc
     1501 aaccgaaatc tctcacacaa tctccggaga ttatattatc atgactatct tttttgacct
     1561 gagcagacgg atgggctatt tcacaattca gacctacatt ccgtgcattc taacagttgt
     1621 tctttcctgg gtgtcttttt ggatcaacaa agatgctgta cctgctagaa catccctggg
     1681 tatcactacg gttttgacta tgacaaccct cagtacaatc gctagaaaat ctttacctaa
     1741 ggtttcttac gtgacagcaa tggatctctt tgtctctgtg tgcttcatct ttgtgttttc
     1801 agctttggtg gagtatggta ccctgcacta ttttgtgagc aaccggaaac caagcaagga
     1861 taaagacaaa aagaagaaaa accctgcccc taccattgat atccgtccca gatcagcaac
     1921 gatccaaatg aacaatgcca cccaccttca agagagggat gaagaatatg gctatgagtg
     1981 tttggatggc aaggactgtg ccagtttctt ttgctgtttt gaagactgcc gaacaggagc
     2041 ctggagacac gggaggatac acattcgcat tgccaaaatg gactcctact ccaggatctt
     2101 tttcccaaca gcttttgcct tgttcaatct ggtttactgg gttggatatc tatacttata
     2161 actcgagtct agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg
     2221 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     2281 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     2341 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     2401 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     2461 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     2521 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     2581 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     2641 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     2701 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     2761 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     2821 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     2881 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     2941 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3001 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3061 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     3121 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     3181 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     3241 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     3301 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     3361 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     3421 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     3481 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     3541 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     3601 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     3661 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     3721 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     3781 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     3841 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     3901 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     3961 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     4021 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     4081 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     4141 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     4201 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     4261 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     4321 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     4381 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     4441 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     4501 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     4561 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     4621 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     4681 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     4741 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     4801 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     4861 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     4921 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     4981 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5041 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     5101 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     5161 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     5221 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     5281 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5341 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     5401 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5461 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     5521 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     5581 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     5641 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5701 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     5761 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     5821 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     5881 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     5941 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     6001 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     6061 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     6121 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     6181 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     6241 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     6301 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     6361 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     6421 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     6481 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6541 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     6601 ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg
     6661 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga
     6721 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgc
//
